Binding Agents

ABSTRACT

Compositions and methods relating to epitopes of sclerostin protein, and sclerostin binding agents, such as antibodies capable of binding to sclerostin, are provided.

RELATED APPLICATIONS

The present application claims benefit of priority from U.S. Provisionalpatent application titled “BINDING AGENTS AND EPITOPES III” Ser. No.60/792,645 filed Apr. 17, 2006, U.S. Provisional Patent Application Ser.No. 60/782,244 filed Mar. 13, 2006, U.S. Provisional Patent ApplicationSer. No. 60/776,847 filed Feb. 24, 2006 and U.S. Provisional PatentApplication Ser. No. 60/677,583 filed May 3, 2005, under 35 U.S.C. §119. The foregoing provisional patent applications are incorporatedherein by reference in their entirety.

TECHNICAL FIELD

The present invention relates generally to epitopes of sclerostinprotein, including human sclerostin protein, and binding agents (such asantibodies) capable of binding to sclerostin or fragments thereof

BACKGROUND OF THE INVENTION

Two or three distinct phases of changes to bone mass occur over the lifeof an individual (see Riggs, West I Med. 154:63-77 (1991)). The firstphase occurs in both men and women and proceeds to attainment of a peakbone mass. This first phase is achieved through linear growth of theendochondral growth plates and radial growth due to a rate of periostealapposition. The second phase begins around age 30 for trabecular bone(flat bones such as the vertebrae and pelvis) and about age 40 forcortical bone (e.g., long bones found in the limbs) and continues to oldage. This phase is characterized by slow bone loss and occurs in bothmen and women. In women, a third phase of bone loss also occurs, mostlikely due to postmenopausal estrogen deficiencies. During this phasealone, women may lose an additional bone mass from the cortical bone andfrom the trabecular compartment (see Riggs, supra).

Loss of bone mineral content can be caused by a wide variety ofconditions and may result in significant medical problems. For example,osteoporosis is a debilitating disease in humans and is characterized bymarked decreases in skeletal bone mass and mineral density, structuraldeterioration of bone, including degradation of bone microarchitectureand corresponding increases in bone fragility (i.e., decreases in bonestrength), and susceptibility fracture in afflicted individuals.Osteoporosis in humans is generally preceded by clinical osteopenia(bone mineral density that is greater than one standard deviation butless than 2.5 standard deviations below the mean value for young adultbone), a condition found in approximately 25 million people in theUnited States. Another 7-8 million patients in the United States havebeen diagnosed with clinical osteoporosis (defined as bone mineralcontent greater than 2.5 standard deviations below that of mature youngadult bone). The frequency of osteoporosis in the human populationincreases with age. Among Caucasians, osteoporosis is predominant inwomen who, in the United States, comprise 80% of the osteoporosispatient pool. The increased fragility and susceptibility to fracture ofskeletal bone in the aged is aggravated by the greater risk ofaccidental falls in this population. Fractured hips, wrists, andvertebrae are among the most common injuries associated withosteoporosis. Hip fractures in particular are extremely uncomfortableand expensive for the patient, and for women, correlate with high ratesof mortality and morbidity.

Although osteoporosis has been regarded as an increase in the risk offracture due to decreased bone mass, few of the presently availabletreatments for skeletal disorders can increase the bone density ofadults, and most of the presently available treatments work primarily byinhibiting further bone resorption rather than stimulating new boneformation. Estrogen is now being prescribed to retard bone loss.However, some controversy exists over whether patients gain anylong-term benefit and whether estrogen has any effect on patients over75 years old. Moreover, use of estrogen is believed to increase the riskof breast and endometrial cancer. Calcitonin, osteocalcin with vitaminK, or high doses of dietary calcium, with or without vitamin D, havealso been suggested for postmenopausal women. High doses of calcium,however, often have undesired gastrointestinal side effects, and serumand urinary calcium levels must be continuously monitored (e.g., Khoslaand Riggs, Mayo Clin. Proc. 70:978982, 1995).

Other current therapeutic approaches to osteoporosis includebisphosphonates (e.g., Fosamax™, Actonel™, Bonviva™, Zometa™,olpadronate, neridronate, skelid, bonefos), parathyroid hormone,calcilytics, calcimimetics (e.g., cinacalcet), statins, anabolicsteroids, lanthanum and strontium salts, and sodium fluoride. Suchtherapeutics, however, are often associated with undesirable sideeffects (see Khosla and Riggs, supra).

Sclerostin, the product of the SOST gene, is absent in sclerosteosis, askeletal disease characterized by bone overgrowth and strong dense bones(Brunkow et al., Am. J. Hum. Genet., 68:577-589, 2001; Balemans et al.,Hum. Mol. Genet., 10:537-543, 2001). The amino acid sequence of humansclerostin is reported by Brunkow et al. ibid and is disclosed herein asSEQ ID NO:1.

BRIEF SUMMARY OF THE INVENTION

Disclosed herein are compositions and methods that can be used toincrease at least one of bone formation, bone mineral density, bonemineral content, bone mass, bone quality and bone strength, and thattherefore may be used to treat a wide variety of conditions in which anincrease in at least one of bone formation, bone mineral density, bonemineral content, bone mass, bone quality and bone strength is desirable.The present invention also offers other related advantages describedherein.

The invention relates to regions (epitopes) of human sclerostinrecognized by the binding agents disclosed herein, methods of usingthese epitopes, and methods of making such epitopes.

The invention also relates to epitopes specific to the region ofsclerostin identified as Loop 2, and binding agents which specificallybind to that region.

The invention also relates to epitopes specific to the cystine-knotregion of sclerostin, and binding agents such as antibodies specificallybinding to that region.

The invention relates to binding agents, such as antibodies, thatspecifically bind to sclerostin. The binding agents can be characterizedby their ability to cross-block the binding of at least one antibodydisclosed herein to sclerostin and/or to be cross-blocked from bindingsclerostin by at least one antibody disclosed herein. The antibodies andother binding agents can also be characterized by their binding patternto human sclerostin peptides in a “human sclerostin peptide epitopecompetition binding assay” as disclosed herein.

The invention relates to binding agents, such as antibodies, that canincrease at least one of bone formation, bone mineral density, bonemineral content, bone mass, bone quality and bone strength in a mammal.

The invention relates to binding agents, such as antibodies, that canblock the inhibitory effect of sclerostin in a cell based mineralizationassay.

The invention further relates to polypeptide constructs comprising two,three, or four polypeptide fragments linked by at least one disulfidebond, representing a core region of the cystine-knot of sclerostin, andantibodies capable of specifically binding thereto.

The invention relates to methods of obtaining epitopes suitable for useas immunogens for generating, in mammals, binding agents, such asantibodies capable of binding specifically to sclerostin; in certainembodiments the binding agents generated are capable of neutralizingsclerostin activity in vivo.

The invention relates to a composition for eliciting an antibodyspecific for sclerostin when the composition is administered to ananimal, the composition comprising a polypeptide having the amino acidsequence of SEQ ID NO:6, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQID NO:66, SEQ ID NO:67, SEQ ID NO:68, or SEQ ID NO:69.

The invention also relates to a composition for eliciting an antibodyspecific for sclerostin when the composition is administered to ananimal, the composition comprising at least one polypeptide consistingessentially of the amino acid sequence of SEQ ID NO:2, SEQ ID NO:3, SEQID NO:4 or SEQ ID NO:5; the composition may comprise at least two or atleast three of the amino acid sequences of SEQ ID NO:2, SEQ ID NO:3, SEQID NO:4 and SEQ ID NO:5, and the composition may comprise all four ofthe amino acid sequences of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 andSEQ ID NO:5.

The invention further relates to a composition for eliciting an antibodyspecific for sclerostin when the composition is administered to ananimal, the composition comprising a polypeptide having the amino acidsequences of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQ ID NO:5,wherein SEQ ID NO:2 and 4 are joined by a disulfide bond at amino acidpositions 57 and 111 with reference to SEQ ID NO:1, and SEQ ID NO:3 and5 are joined by at least one of (a) a disulfide bond at amino acidpositions 82 and 142 with reference to SEQ ID NO:1, and (b) a disulfidebond at amino acid positions 86 and 144 with reference to SEQ ID NO:1;the polypeptide may retain the tertiary structure of the correspondingpolypeptide region of human sclerostin of SEQ ID NO:1.

The invention also relates to polypeptide T20.6 consisting essentiallyof a multiply truncated human sclerostin protein of SEQ ID NO:1, whereinamino acids 1-50, 65-72, 91-100, 118-137, and 150-190 of SEQ ID NO:1 areabsent from the polypeptide; this polypeptide may be obtained by trypticdigestion of human sclerostin, and the protein may be isolated by HPLCfractionation.

The invention further relates to immunogenic portion T20.6 of humansclerostin comprising amino acids 51-64, 73-90, 101-117, and 138-149 ofSEQ ID NO:1, wherein the immunogenic portion comprises at least one of:

(a) a disulfide bond between amino acids 57 and 111;

(b) a disulfide bond between amino acids 82 and 142; and

(c) a disulfide bond between amino acids 86 and 144;

the immunogenic portion may have at least two of these disulfide bonds;and the immunogenic portion may have all three disulfide bonds.

The invention further relates to an immunogenic portion T20.6 derivativeof human sclerostin comprising amino acids 57-64, 73-86, 111-117, and138-144 of SEQ ID NO:1, wherein the immunogenic portion comprises atleast one of:

(a) a disulfide bond between amino acids 57 and 111;

(b) a disulfide bond between amino acids 82 and 142; and

(c) a disulfide bond between amino acids 86 and 144;

the immunogenic portion may have at least two of these disulfide bonds;and the immunogenic portion may have all three disulfide bonds.

The invention yet further relates to a polypeptide consistingessentially of a human sclerostin protein of SEQ ID NO:1 truncated atthe C-terminal and N-terminal ends, wherein amino acids 1-85 and 112-190of SEQ ID NO:1 are absent from the polypeptide.

The invention also relates to an immunogenic portion of humansclerostin, comprising amino acids 86-111 of SEQ ID NO:1; theimmunogenic portion may consist essentially of contiguous amino acidsCGPARLLPNAIGRGKWWRPSGPDFRC (SEQ ID NO:6).

The invention further relates to an immunogenic portion of ratsclerostin, comprising amino acids 92-109 of SEQ ID NO:98; theimmunogenic portion may consist essentially of contiguous amino acidsPNAIGRVKWWRPNGPDFR (SEQ ID NO:96).

The invention still further relates to an immunogenic portion of ratsclerostin, comprising amino acids 99-120 of SEQ ID NO:98; theimmunogenic portion may consist essentially of contiguous amino acidsKWWRPNGPDFRCIPDRYRAQRV (SEQ ID NO:97).

The invention relates to a method of producing an immunogenic portion ofhuman sclerostin, comprising the steps of:

-   -   (a) treating human sclerostin to achieve complete tryptic        digestion;    -   (b) collecting the tryptic digest sample having average        molecular weight of 7,122.0 Daltons (theoretical mass 7121.5        Daltons) or retention time of about 20.6 minutes as determined        by elution from a reverse-phase HPLC column with linear gradient        from 0.05% trifluoroacetic acid to 90% acetonitrile in 0.05% TFA        at a flow rate of 0.2 ml/min; and    -   (c) purifying the immunogenic portion.

The invention relates to a method of generating an antibody capable ofspecifically binding to sclerostin, comprising:

-   -   (a) immunizing an animal with a composition comprising a        polypeptide of SEQ ID NO:6, SEQ ID NO:63, SEQ ID NO:64, SEQ ID        NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69,        SEQ ID NO:96, or SEQ ID NO:97;    -   (b) collecting sera from the animal; and    -   (c) isolating from the sera an antibody capable of specifically        binding to sclerostin.

The invention also relates to a method of generating an antibody capableof specifically binding to sclerostin, the method comprising:

-   -   (a) immunizing an animal with a composition comprising        polypeptide T20.6 or a derivative of T20.6;    -   (b) collecting sera from the animal; and    -   (c) isolating from the sera an antibody capable of specifically        binding to sclerostin.

The invention further relates to a method of detecting ananti-sclerostin antibody in a biological sample, comprising the steps of

-   -   (a) contacting the biological sample with a polypeptide        consisting essentially of SEQ ID NO:6, SEQ ID NO:63, SEQ ID        NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68,        SEQ ID NO:69, SEQ ID NO:96, or SEQ ID NO:97 under conditions        allowing a complex to form between the antibody and the        polypeptide; and    -   (b) detecting the presence or absence of the complex,        wherein the presence of the complex indicates that the        biological sample contains an anti-sclerostin antibody.

The invention also relates to a method of detecting an anti-sclerostinantibody in a biological sample, comprising the steps of

-   -   (a) contacting the biological sample with polypeptide T20.6 or a        derivative of T20.6 under conditions allowing a complex to form        between the antibody and the polypeptide; and    -   (b) detecting the presence or absence of the complex,        wherein the presence of the complex indicates that the        biological sample contains an anti-sclerostin antibody.

The invention further relates to a sclerostin binding agent, such as anantibody, that cross-blocks the binding of at least one of antibodiesAb-A, Ab-B, Ab-C, or Ab-D to a sclerostin protein. The sclerostinbinding agent may also be cross-blocked from binding to sclerostin by atleast one of antibodies Ab-A, Ab-B, Ab-C, or Ab-D. The isolatedantibody, or an antigen-binding fragment thereof, may be a polyclonalantibody, a monoclonal antibody, a humanized antibody, a human antibody,a chimeric antibody or the like.

The invention further relates to a sclerostin binding agent, such as anantibody, that is cross-blocked from binding to sclerostin by at leastone of antibodies Ab-A, Ab-B, Ab-C, or Ab-D. The isolated antibody, oran antigen-binding fragment thereof, may be a polyclonal antibody, amonoclonal antibody, a humanized antibody, a human antibody, a chimericantibody or the like.

The invention further relates to a sclerostin binding agent, such as anisolated antibody, that cross-blocks the binding of at least one ofantibodies 1-24 (Ab-1 to Ab-24) to a sclerostin protein. The sclerostinbinding agent may also be cross-blocked from binding to sclerostin by atleast one of antibodies 1-24 (Ab-1 to Ab-24). The isolated antibody, oran antigen-binding fragment thereof, may be a polyclonal antibody, amonoclonal antibody, a humanized antibody, a human antibody, or achimeric antibody.

The invention further relates to a sclerostin binding agent, such as anisolated antibody, that is cross-blocked from binding to sclerostin byat least one of antibodies 1-24 (Ab-1 to Ab-24); the isolated antibody,or an antigen-binding fragment thereof, may be a polyclonal antibody, amonoclonal antibody, a humanized antibody, a human antibody, or achimeric antibody.

The invention further relates to a binding agent, such as an isolatedantibody that exhibits a similar binding pattern to human sclerostinpeptides in a “human sclerostin peptide epitope competition bindingassay” as that exhibited by at least one of the antibodies Ab-A, Ab-B,Ab-C or Ab-D; the isolated antibody, or an antigen-binding fragmentthereof, may be a polyclonal antibody, a monoclonal antibody, ahumanized antibody, a human antibody, or a chimeric antibody.

The invention still further relates to a method for treating a bonedisorder associated with at least one of low bone formation, low bonemineral density, low bone mineral content, low bone mass, low bonequality and low bone strength in a mammalian subject which comprisesproviding to a subject in need of such treatment an amount of ananti-sclerostin binding agent sufficient to increase at least one ofbone formation, bone mineral density, bone mineral content, bone mass,bone quality and bone strength wherein the anti-sclerostin binding agentcomprises an antibody, or sclerostin-binding fragment thereof.

The invention also relates to an isolated sclerostin polypeptide orfragments thereof, wherein the polypeptide contains 6 conserved cysteineresidues and the fragments thereof comprise from 7 to 14 amino acids ofSEQ ID NO:2; 8 to 17 amino acids of SEQ ID NO:3; 8 to 18 residues of SEQID NO:4; and 6 to 12 residues of SEQ ID NO:5, and the polypeptide orfragments thereof are stabilized by disulfide bonds between SEQ ID NO:2and 4, and between SEQ ID NO:3 and 5; the polypeptide or fragments maycomprise 10-14 amino acids of SEQ ID NO:2; 14 to 17 amino acids of SEQID NO:3; 13 to 18 amino acids of SEQ ID NO:4; and 8 to 12 residues ofSEQ ID NO:5; and the polypeptide or fragments may comprise SEQ ID NO:2,SEQ ID NO:3, SEQ ID NO:4, and SEQ ID NO:5.

Provided herein are antibodies that specifically bind to humansclerostin. The antibodies are characterized by their ability tocross-block the binding of at least one antibody disclosed herein tohuman sclerostin and/or to be cross-blocked from binding humansclerostin by at least one antibody disclosed herein.

Also provided is an isolated antibody, or an antigen-binding fragmentthereof, that can increase at least one of bone formation, bone mineraldensity, bone mineral content, bone mass, bone quality and bone strengthin a mammal.

Also provided in an isolated antibody, or an antigen-binding fragmentthereof, that can block the inhibitory effect of sclerostin in a cellbased mineralization assay.

Also provided is a binding agent, such as an antibody, that specificallybinds to human sclerostin and has at least one CDR sequence selectedfrom SEQ ID NOs: 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 78, 79, 80, 81, 99, 100, 101,102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115,116, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249,250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263,264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291,292, 293, 294, 295, 296, 297, 298, 351, 352, 353, 358, 359, and 360, andvariants thereof, wherein the antibody or antigen-binding fragmentthereof neutralizes sclerostin.

Also provided is a binding agent, such as an antibody, that specificallybinds to human sclerostin and has at least one CDR sequence selectedfrom SEQ ID NOs:39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 78, 79, 80, 81, 99, 100, 101,102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115,116, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249,250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263,264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291,292, 293, 294, 295, 296, 297, 298, 351, 352, 353, 358, 359, and 360, andvariants thereof.

Also provided are regions of human sclerostin which are important forthe in vivo activity of the protein.

These and other aspects of the present invention will become apparentupon reference to the following detailed description and attacheddrawings. All references disclosed herein are hereby incorporated byreference in their entireties as if each was incorporated individually.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts the amino acid sequences of the mature form (signalpeptides cleaved off) of the light chain (FIG. 1A) (SEQ ID NO:23) andheavy chain (FIG. 1B) (SEQ ID NO:27) for the anti-human sclerostin andanti-mouse sclerostin antibody Ab-A.

FIG. 2 depicts the amino acid sequences of the mature form (signalpeptides cleaved off) of the light chain (FIG. 2A) (SEQ ID NO:31) andheavy chain (FIG. 2B) (SEQ ID NO:35) for the anti-human sclerostin andanti-mouse sclerostin antibody Ab-B.

FIG. 3 depicts the amino acid sequences of the mature form (signalpeptides cleaved oft) of the light chain (FIG. 3A) (SEQ ID NO:15) andheavy chain (FIG. 3B) (SEQ ID NO:19) for the anti-human sclerostin andanti-mouse sclerostin antibody Ab-C.

FIG. 4 depicts the amino acid sequences of the mature form (signalpeptides cleaved off) of the light chain (FIG. 4A) (SEQ ID NO:7) andheavy chain (FIG. 4B) (SEQ ID NO:11) for the anti-human sclerostin andanti-mouse sclerostin antibody Ab-D.

FIG. 5 depicts bone mineral density in mice measured at two skeletalsites (lumbar vertebrae and tibial metaphysis) after 3 weeks oftreatment with vehicle, PTH (1-34), Ab-A or Ab-B.

FIG. 6 shows bone mineral density in mice measured at two skeletal sites(lumbar vertebrae and tibial metaphysis) after 2 weeks of treatment withvehicle, PTH (1-34) or Ab-C.

FIG. 7 depicts bone mineral density in mice measured at two skeletalsites (lumbar vertebrae and tibial metaphysis) after 3 weeks oftreatment with vehicle or Ab-D.

FIG. 8 depicts the amino acid sequence of the mature form (signalpeptide cleaved oft) of human sclerostin (SEQ ID NO:1). Also depicted isthe nucleotide sequence of the human sclerostin coding region thatencodes the mature form of human sclerostin. The eight cysteines arenumbered C1 through C8. The cystine-knot is formed by three disulfidebonds (C1-C5; C3-C7; C4-C8). C2 and C6 also form a disulfide bond,however this disulfide is not part of the cystine-knot.

FIG. 9 depicts a schematic of the basic structure of human sclerostin.There is an N-terminal arm (from the first Q to C1) and a C-terminal arm(from C8 to the terminal Y). In between these arms there is thecystine-knot structure (formed by three disulfides: C1-C5; C3-C7; C4-C8)and three loops which are designated Loop1, Loop 2 and Loop 3. Thedistal regions of Loop 1 and Loop 3 are linked by the C2-C6 disulfide.Potential trypsin cleavage sites are indicated (arginine=R andlysine=K). Some of the potential AspN cleavage sites are indicated (onlyaspartic acid (D) residues are shown).

FIG. 10 depicts the HPLC peptide maps of human sclerostin afterdigestion with either trypsin or AspN. The human sclerostin peptidesgenerated by trypsin digestion are indicated (T19.2, T20, T20.6 andT21-22) as are the human sclerostin peptides generated by AspN digestion(AspN14.6, AspN18.6 and AspN22.7-23.5).

FIG. 11 depicts sequence and mass information for the isolated humansclerostin disulfide linked peptides generated by trypsin digestion.Seq. pos.=sequence position. Obs.=observed. Observed mass was determinedby ESI-LC-MS analysis.

FIG. 12 depicts sequence and mass information for the isolated humansclerostin peptides generated by AspN digestion. The AspN22.7-23.5peptide contains the 4 disulfide bonds. Seq. pos.=sequence position.Obs.=observed. Observed mass was determined by ESI-LC-MS analysis.

FIG. 13 shows a linear schematic of four human sclerostin peptides(T19.2, T20, T20.6 and T21-22) generated by trypsin digestion.

FIG. 14 shows a linear schematic of five human sclerostin peptides(AspN14.6, AspN18.6 and AspN22.7-23.5) generated by AspN digestion. TheAspN14.6 HPLC peak is composed of three peptides not linked by anydisulfide bonds.

FIG. 15 shows the resonance unit (Ru) signal from the Biacore-based“human sclerostin peptide epitope competition binding assay.” RelativeMab binding to various human sclerostin-peptides (in solution) versusMab binding to intact mature form human sclerostin (immobilized onBiacore chip) was assessed. Data shown is for Ab-A. Human sclerostinpeptides used were T19.2, T20, T20.6, T21-22, AspN14.6, AspN18.6 andAspN22.7-23.5.

FIG. 16 shows the resonance unit (Ru) signal from the Biacore-based“human sclerostin peptide epitope competition binding assay.” RelativeMab binding to various human sclerostin-peptides (in solution) versusMab binding to intact mature form human sclerostin (immobilized onBiacore chip) was assessed. Data shown is for Ab-B. Human sclerostinpeptides used were T19.2, T20, T20.6, T21-22, AspN14.6, AspN18.6 andAspN22.7-23.5.

FIG. 17 shows the resonance unit (Ru) signal from the Biacore-based“human sclerostin peptide epitope competition binding assay.” RelativeMab binding to various human sclerostin-peptides (in solution) versusMab binding to intact mature form human sclerostin (immobilized onBiacore chip) was assessed. Data shown is for Ab-C. Human sclerostinpeptides used were T19.2, T20, T20.6, T21-22, AspN14.6, AspN18.6 andAspN22.7-23.5.

FIG. 18 shows the resonance unit (Ru) signal from Biacore-based “humansclerostin peptide epitope competition binding assay.” Relative Mabbinding to various human sclerostin-peptides (in solution) versus Mabbinding to intact mature form human sclerostin (immobilized on Biacorechip) was assessed. Data shown is for Ab-D. Human sclerostin peptidesused were T19.2, T20, T20.6, T21-22, AspN14.6, AspN18.6 andAspN22.7-23.5.

FIG. 19 shows two Mab binding epitopes of human sclerostin. FIG. 19Ashows sequence of the Loop 2 epitope for binding of Ab-A and Ab-B tohuman sclerostin (SEQ ID NO:6). FIG. 19B shows sequence, disulfidebonding and schematic of the T20.6 epitope for binding of Ab-C and Ab-Dto human sclerostin (SEQ ID NO:2-5).

FIG. 20 depicts the HPLC peptide maps of human sclerostin afterdigestion with trypsin. FIG. 20A shows digestion of the human sclerostinAb-D complex. FIG. 20B shows digestion of human sclerostin alone. TheT19.2, T20, T20.6 and T21-22 peptide peaks are indicated.

FIG. 21 shows the sequence, disulfide bonding and schematic of the“T20.6 derivative 1 (cystine-knot+4 arms)” epitope for binding of Ab-Dto human sclerostin. (SEQ ID NO:70-73).

FIG. 22 shows results from the MC3T3-E1-BF osteoblast cell linemineralization assay used for identifying anti-sclerostin neutralizingMabs. Mouse sclerostin (Scl) was used at 1 μg/ml. Monoclonal antibodieswere used at 10 and 5 μg/ml. Extent of mineralization (various types ofinsoluble calcium phosphate) was quantitated by measuring calcium.

FIG. 23 depicts results from the MC3T3-E1-BF osteoblast cell linemineralization assay used for identifying anti-sclerostin neutralizingMabs. Human sclerostin (Scl) was used at 1 μg/ml. Monoclonal antibodieswere used at 8 and 4 μg/ml. Extent of mineralization (various types ofinsoluble calcium phosphate) was quantitated by measuring calcium.

FIG. 24 shows results from the MC3T3-E1-BF osteoblast cell linemineralization assay used for identifying anti-sclerostin neutralizingMabs. Human sclerostin (Scl) was used at 1 μg/ml. Monoclonal antibodieswere used at 10 μg/ml. Extent of mineralization (various types ofinsoluble calcium phosphate) was quantitated by measuring calcium.

FIG. 25 depicts results from an inflammation-induced bone loss SCIDmouse model. Ab-A treatment protected mice from inflammation-relatedbone loss associated with colitis when measured as total bone mineraldensity (FIG. 25A), vertebral bone density (FIG. 25B), and femur bonedensity (FIG. 25C).

DETAILED DESCRIPTION

The present invention relates to regions of the human sclerostin proteinthat contain epitopes recognized by antibodies that also bind tofull-length sclerostin, and methods of making and using these epitopes.The invention also provides binding agents (such as antibodies) thatspecifically bind to sclerostin or portions of sclerostin, and methodsfor using such binding agents. The binding agents are useful to block orimpair binding of human sclerostin to one or more ligand.

Recombinant human sclerostin/SOST is commercially available from R&DSystems (Minneapolis, Minn., USA; 2006 cat #1406-ST-025). Additionally,recombinant mouse sclerostin/SOST is commercially available from R&DSystems (Minneapolis, Minn., USA; 2006 cat #1589-ST-025). Research gradesclerostin binding monoclonal antibodies are commercially available fromR&D Systems (Minneapolis, Minn., USA; mouse monoclonal: 2006 cat #MAB1406; rat monoclonal: 2006 cat # MAB1589). U.S. Pat. Nos. 6,395,511and 6,803,453, and U.S. Patent Publications 20040009535 and 20050106683refer to anti-sclerostin antibodies generally.

As used herein, the term human sclerostin is intended to include theprotein of SEQ ID NO:1 and allelic variants thereof. Sclerostin can bepurified from 293T host cells that have been transfected by a geneencoding sclerostin by elution of filtered supernatant of host cellculture fluid using a Heparin HP column, using a salt gradient. Thepreparation and further purification using cation exchangechromatography are described in Examples 1 and 2.

Binding agents of the invention are preferably antibodies, as definedherein. The term “antibody” refers to an intact antibody, or a bindingfragment thereof. An antibody may comprise a complete antibody molecule(including polyclonal, monoclonal, chimeric, humanized, or humanversions having full length heavy and/or light chains), or comprise anantigen binding fragment thereof. Antibody fragments include F(ab′)₂,Fab, Fab′, Fv, Fc, and Fd fragments, and can be incorporated into singledomain antibodies, single-chain antibodies, maxibodies, minibodies,intrabodies, diabodies, triabodies, tetrabodies, v-NAR and bis-scFv (Seee.g., Hollinger and Hudson, 2005, Nature Biotechnology, 23, 9,1126-1136). Antibody polypeptides are also disclosed in U.S. Pat. No.6,703,199, including fibronectin polypeptide monobodies. Other antibodypolypeptides are disclosed in U.S. Patent Publication 2005/0238646,which are single-chain polypeptides.

Antigen binding fragments derived from an antibody can be obtained, forexample, by proteolytic hydrolysis of the antibody, for example, pepsinor papain digestion of whole antibodies according to conventionalmethods. By way of example, antibody fragments can be produced byenzymatic cleavage of antibodies with pepsin to provide a 5S fragmenttermed F(ab′)₂. This fragment can be further cleaved using a thiolreducing agent to produce 3.5S Fab′ monovalent fragments. Optionally,the cleavage reaction can be performed using a blocking group for thesulfhydryl groups that result from cleavage of disulfide linkages. As analternative, an enzymatic cleavage using papain produces two monovalentFab fragments and an Fc fragment directly. These methods are described,for example, by Goldenberg, U.S. Pat. No. 4,331,647, Nisonoff et al.,Arch. Biochem. Biophys. 89:230, 1960; Porter, Biochem. J. 73:119, 1959;Edelman et al., in Methods in Enzymology 1:422 (Academic Press 1967);and by Andrews, S. M. and Titus, J. A. in Current Protocols inImmunology (Coligan J. E., et al., eds), John Wiley & Sons, New York(2003). pages 2.8.1-2.8.10 and 2.10A.1-2.10A.5. Other methods forcleaving antibodies, such as separating heavy chains to form monovalentlight-heavy chain fragments (Fd), further cleaving of fragments, orother enzymatic, chemical, or genetic techniques may also be used, solong as the fragments bind to the antigen that is recognized by theintact antibody.

An antibody fragment may also be any synthetic or genetically engineeredprotein. For example, antibody fragments include isolated fragmentsconsisting of the light chain variable region, “Fv” fragments consistingof the variable regions of the heavy and light chains, recombinantsingle chain polypeptide molecules in which light and heavy variableregions are connected by a peptide linker (scFv proteins).

Another form of an antibody fragment is a peptide comprising one or morecomplementarity determining regions (CDRs) of an antibody. CDRs (alsotermed “minimal recognition units”, or “hypervariable region”) can beobtained by constructing polynucleotides that encode the CDR ofinterest. Such polynucleotides are prepared, for example, by using thepolymerase chain reaction to synthesize the variable region using mRNAof antibody-producing cells as a template (see, for example, Larrick etal., Methods: A Companion to Methods in Enzymology 2:106, 1991;Courtenay-Luck, “Genetic Manipulation of Monoclonal Antibodies,” inMonoclonal Antibodies: Production, Engineering and Clinical Application,Ritter et al. (eds.), page 166 (Cambridge University Press 1995); andWard et al., “Genetic Manipulation and Expression of Antibodies,” inMonoclonal Antibodies: Principles and Applications, Birch et al.,(eds.), page 137 (Wiley-Liss, Inc. 1995)).

Thus, in one embodiment, the binding agent comprises at least one CDR asdescribed herein. The binding agent may comprise at least two, three,four, five or six CDR's as described herein. The binding agent furthermay comprise at least one variable region domain of an antibodydescribed herein. The variable region domain may be of any size or aminoacid composition and will generally comprise at least one CDR sequenceresponsible for binding to human sclerostin, for example CDR-H1, CDR-H2,CDR-H3 and/or the light chain CDRs specifically described herein andwhich is adjacent to or in frame with one or more framework sequences.In general terms, the variable (V) region domain may be any suitablearrangement of immunoglobulin heavy (V_(H)) and/or light (V_(L)) chainvariable domains. Thus, for example, the V region domain may bemonomeric and be a V_(H) or V_(L) domain, which is capable ofindependently binding human sclerostin with an affinity at least equalto 1×10⁻⁷M or less as described below. Alternatively, the V regiondomain may be dimeric and contain V_(H)—V_(H), V_(H)—V_(L), orV_(L)—V_(L), dimers. The V region dimer comprises at least one V_(H) andat least one V_(L) chain that may be non-covalently associated(hereinafter referred to as F_(V)). If desired, the chains may becovalently coupled either directly, for example via a disulfide bondbetween the two variable domains, or through a linker, for example apeptide linker, to form a single chain Fv (scF_(V)).

The variable region domain may be any naturally occurring variabledomain or an engineered version thereof. By engineered version is meanta variable region domain that has been created using recombinant DNAengineering techniques. Such engineered versions include those created,for example, from a specific antibody variable region by insertions,deletions, or changes in or to the amino acid sequences of the specificantibody. Particular examples include engineered variable region domainscontaining at least one CDR and optionally one or more framework aminoacids from a first antibody and the remainder of the variable regiondomain from a second antibody.

The variable region domain may be covalently attached at a C-terminalamino acid to at least one other antibody domain or a fragment thereof.Thus, for example, a VH domain that is present in the variable regiondomain may be linked to an immunoglobulin CH1 domain, or a fragmentthereof. Similarly a V_(L) domain may be linked to a C_(K) domain or afragment thereof. In this way, for example, the antibody may be a Fabfragment wherein the antigen binding domain contains associated V_(H)and V_(L) domains covalently linked at their C-termini to a CH1 andC_(K) domain, respectively. The CH1 domain may be extended with furtheramino acids, for example to provide a hinge region or a portion of ahinge region domain as found in a Fab′ fragment, or to provide furtherdomains, such as antibody CH2 and CH3 domains.

As described herein, binding agents comprise at least one of these CDRs.For example, one or more CDR may be incorporated into known antibodyframework regions (IgG1, IgG2, etc.), or conjugated to a suitablevehicle to enhance the half-life thereof. Suitable vehicles include, butare not limited to Fc, polyethylene glycol (PEG), albumin, transferrin,and the like. These and other suitable vehicles are known in the art.Such conjugated CDR peptides may be in monomeric, dimeric, tetrameric,or other form. In one embodiment, one or more water-soluble polymer isbonded at one or more specific position, for example at the aminoterminus, of a binding agent.

In certain preferred embodiments, a binding agent comprises one or morewater soluble polymer attachments, including, but not limited to,polyethylene glycol, polyoxyethylene glycol, or polypropylene glycol.See, e.g., U.S. Pat. Nos. 4,640,835, 4,496,689, 4,301,144, 4,670,417,4,791,192 and 4,179,337. In certain embodiments, a derivative bindingagent comprises one or more of monomethoxy-polyethylene glycol, dextran,cellulose, or other carbohydrate based polymers, poly-(N-vinylpyrrolidone)-polyethylene glycol, propylene glycol homopolymers, apolypropylene oxide/ethylene oxide co-polymer, polyoxyethylated polyols(e.g., glycerol) and polyvinyl alcohol, as well as mixtures of suchpolymers. In certain embodiments, one or more water-soluble polymer israndomly attached to one or more side chains. In certain embodiments,PEG can act to improve the therapeutic capacity for a binding agent,such as an antibody. Certain such methods are discussed, for example, inU.S. Pat. No. 6,133,426, which is hereby incorporated by reference forany purpose.

It will be appreciated that a binding agent of the present invention mayhave at least one amino acid substitution, providing that the bindingagent retains binding specificity. Therefore, modifications to thebinding agent structures are encompassed within the scope of theinvention. These may include amino acid substitutions, which may beconservative or non-conservative, that do not destroy the sclerostinbinding capability of a binding agent. Conservative amino acidsubstitutions may encompass non-naturally occurring amino acid residues,which are typically incorporated by chemical peptide synthesis ratherthan by synthesis in biological systems. These include peptidomimeticsand other reversed or inverted forms of amino acid moieties. Aconservative amino acid substitution may also involve a substitution ofa native amino acid residue with a normative residue such that there islittle or no effect on the polarity or charge of the amino acid residueat that position.

Non-conservative substitutions may involve the exchange of a member ofone class of amino acids or amino acid mimetics for a member fromanother class with different physical properties (e.g. size, polarity,hydrophobicity, charge). Such substituted residues may be introducedinto regions of the human antibody that are homologous with non-humanantibodies, or into the non-homologous regions of the molecule.

Moreover, one skilled in the art may generate test variants containing asingle amino acid substitution at each desired amino acid residue. Thevariants can then be screened using activity assays known to thoseskilled in the art. Such variants could be used to gather informationabout suitable variants. For example, if one discovered that a change toa particular amino acid residue resulted in destroyed, undesirablyreduced, or unsuitable activity, variants with such a change may beavoided. In other words, based on information gathered from such routineexperiments, one skilled in the art can readily determine the aminoacids where further substitutions should be avoided either alone or incombination with other mutations.

A skilled artisan will be able to determine suitable variants of thepolypeptide as set forth herein using well-known techniques. In certainembodiments, one skilled in the art may identify suitable areas of themolecule that may be changed without destroying activity by targetingregions not believed to be important for activity. In certainembodiments, one can identify residues and portions of the moleculesthat are conserved among similar polypeptides. In certain embodiments,even areas that may be important for biological activity or forstructure may be subject to conservative amino acid substitutionswithout destroying the biological activity or without adverselyaffecting the polypeptide structure.

Additionally, one skilled in the art can review structure-functionstudies identifying residues in similar polypeptides that are importantfor activity or structure. In view of such a comparison, one can predictthe importance of amino acid residues in a protein that correspond toamino acid residues which are important for activity or structure insimilar proteins. One skilled in the art may opt for chemically similaramino acid substitutions for such predicted important amino acidresidues.

One skilled in the art can also analyze the three-dimensional structureand amino acid sequence in relation to that structure in similarpolypeptides. In view of such information, one skilled in the art maypredict the alignment of amino acid residues of an antibody with respectto its three dimensional structure. In certain embodiments, one skilledin the art may choose not to make radical changes to amino acid residuespredicted to be on the surface of the protein, since such residues maybe involved in important interactions with other molecules.

A number of scientific publications have been devoted to the predictionof secondary structure. See Moult J., Curr. Op. in Biotech.,7(4):422-427 (1996), Chou et al., Biochemistry, 13(2):222-245 (1974);Chou et al., Biochemistry, 113(2):211-222 (1974); Chou et al., Adv.Enzymol. Relat. Areas Mol. Biol., 47:45-148 (1978); Chou et al., Ann.Rev. Biochem., 47:251-276 and Chou et al., Biophys. J., 26:367-384(1979). Moreover, computer programs are currently available to assistwith predicting secondary structure. One method of predicting secondarystructure is based upon homology modeling. For example, two polypeptidesor proteins which have a sequence identity of greater than 30%, orsimilarity greater than 40% often have similar structural topologies.The recent growth of the protein structural database (PDB) has providedenhanced predictability of secondary structure, including the potentialnumber of folds within a polypeptide's or protein's structure. See Holmet al., Nucl. Acid. Res., 27(1):244-247 (1999). It has been suggested(Brenner et al., Curr. Op. Struct. Biol., 7(3):369-376 (1997)) thatthere are a limited number of folds in a given polypeptide or proteinand that once a critical number of structures have been resolved,structural prediction will become dramatically more accurate.

Additional methods of predicting secondary structure include “threading”(Jones, D., Curr. Opin. Struct. Biol., 7(3):377-87 (1997); Sippl et al.,Structure, 4(1):15-19 (1996)), “profile analysis” (Bowie et al.,Science, 253:164-170 (1991); Gribskov et al., Meth. Enzym., 183:146-159(1990); Gribskov et al., Proc. Nat. Acad. Sci., 84(13):4355-4358(1987)), and “evolutionary linkage” (See Holm, supra (1999), andBrenner, supra (1997)).

In certain embodiments, variants of binding agents include glycosylationvariants wherein the number and/or type of glycosylation site has beenaltered compared to the amino acid sequences of a parent polypeptide. Incertain embodiments, variants comprise a greater or a lesser number ofN-linked glycosylation sites than the native protein. An N-linkedglycosylation site is characterized by the sequence: Asn-X-Ser orAsn-X-Thr, wherein the amino acid residue designated as X may be anyamino acid residue except proline. The substitution of amino acidresidues to create this sequence provides a potential new site for theaddition of an N-linked carbohydrate chain. Alternatively, substitutionswhich eliminate this sequence will remove an existing N-linkedcarbohydrate chain. Also provided is a rearrangement of N-linkedcarbohydrate chains wherein one or more N-linked glycosylation sites(typically those that are naturally occurring) are eliminated and one ormore new N-linked sites are created. Additional preferred antibodyvariants include cysteine variants wherein one or more cysteine residuesare deleted from or substituted for another amino acid (e.g., serine) ascompared to the parent amino acid sequence. Cysteine variants may beuseful when antibodies must be refolded into a biologically activeconformation such as after the isolation of insoluble inclusion bodies.Cysteine variants generally have fewer cysteine residues than the nativeprotein, and typically have an even number to minimize interactionsresulting from unpaired cysteines.

Desired amino acid substitutions (whether conservative ornon-conservative) can be determined by those skilled in the art at thetime such substitutions are desired. In certain embodiments, amino acidsubstitutions can be used to identify important residues of antibodiesto sclerostin, or to increase or decrease the affinity of the antibodiesto sclerostin described herein.

According to certain embodiments, preferred amino acid substitutions arethose which: (1) reduce susceptibility to proteolysis, (2) reducesusceptibility to oxidation, (3) alter binding affinity for formingprotein complexes, (4) alter binding affinities, and/or (4) confer ormodify other physiochemical or functional properties on suchpolypeptides. According to certain embodiments, single or multiple aminoacid substitutions (in certain embodiments, conservative amino acidsubstitutions) may be made in the naturally-occurring sequence (incertain embodiments, in the portion of the polypeptide outside thedomain(s) forming intermolecular contacts). In certain embodiments, aconservative amino acid substitution typically may not substantiallychange the structural characteristics of the parent sequence (e.g., areplacement amino acid should not tend to break a helix that occurs inthe parent sequence, or disrupt other types of secondary structure thatcharacterizes the parent sequence). Examples of art-recognizedpolypeptide secondary and tertiary structures are described in Proteins,Structures and Molecular Principles (Creighton, Ed., W. H. Freeman andCompany, New York (1984)); Introduction to Protein Structure (C. Brandenand J. Tooze, eds., Garland Publishing, New York, N.Y. (1991)); andThornton et al. Nature 354:105 (1991), which are each incorporatedherein by reference.

In certain embodiments, binding agents of the invention may bechemically bonded with polymers, lipids, or other moieties.

The binding agents may comprise at least one of the CDRs describedherein incorporated into a biocompatible framework structure. In oneexample, the biocompatible framework structure comprises a polypeptideor portion thereof that is sufficient to form a conformationally stablestructural support, or framework, or scaffold, which is able to displayone or more sequences of amino acids that bind to an antigen (e.g.,CDRs, a variable region, etc.) in a localized surface region. Suchstructures can be a naturally occurring polypeptide or polypeptide“fold” (a structural motif), or can have one or more modifications, suchas additions, deletions or substitutions of amino acids, relative to anaturally occurring polypeptide or fold. These scaffolds can be derivedfrom a polypeptide of any species (or of more than one species), such asa human, other mammal, other vertebrate, invertebrate, plant, bacteriaor virus.

Typically the biocompatible framework structures are based on proteinscaffolds or skeletons other than immunoglobulin domains. For example,those based on fibronectin, ankyrin, lipocalin, neocarzinostain,cytochrome b, CP1 zinc finger, PST1, coiled coil, LACI-D1, Z domain andtendramisat domains may be used (See e.g., Nygren and Uhlen, 1997,Current Opinion in Structural Biology, 7, 463-469).

In preferred embodiments, it will be appreciated that the binding agentsof the invention include the humanized antibodies described herein.Humanized antibodies such as those described herein can be producedusing techniques known to those skilled in the art (Zhang, W., et al.,Molecular Immunology. 42(12):1445-1451, 2005; Hwang W. et al., Methods.36(1):35-42, 2005; Dall'Acqua W F, et al., Methods 36(1):43-60, 2005;and Clark, M., Immunology Today. 21(8):397-402, 2000).

Additionally, one skilled in the art will recognize that suitablebinding agents include portions of these antibodies, such as one or moreof CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 as specificallydisclosed herein. At least one of the regions of CDR-H1, CDR-H2, CDR-H3,CDR-L1, CDR-L2 and CDR-L3 may have at least one amino acid substitution,provided that the binding agent retains the binding specificity of thenon-substituted CDR. The non-CDR portion of the binding agent may be anon-protein molecule, wherein the binding agent cross-blocks the bindingof an antibody disclosed herein to sclerostin and/or neutralizessclerostin. The non-CDR portion of the binding agent may be anon-protein molecule in which the binding agent exhibits a similarbinding pattern to human sclerostin peptides in a “human sclerostinpeptide epitope competition binding assay” as that exhibited by at leastone of antibodies Ab-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5,Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16,Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, Ab-22, Ab-23, and Ab-24, and/orneutralizes sclerostin. The non-CDR portion of the binding agent may becomposed of amino acids, wherein the binding agent is a recombinantbinding protein or a synthetic peptide, and the recombinant bindingprotein cross-blocks the binding of an antibody disclosed herein tosclerostin and/or neutralizes sclerostin. The non-CDR portion of thebinding agent may be composed of amino acids, wherein the binding agentis a recombinant binding protein, and the recombinant binding proteinexhibits a similar binding pattern to human sclerostin peptides in thehuman sclerostin peptide epitope competition binding assay (describedhereinbelow) as that exhibited by at least one of the antibodies Ab-A,Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9,Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19,Ab-20, Ab-21, Ab-22, Ab-23, and Ab-24, and/or neutralizes sclerostin.

Where an antibody comprises one or more of CDR-H1, CDR-H2, CDR-H3,CDR-L1, CDR-L2 and CDR-L3 as described above, it may be obtained byexpression from a host cell containing DNA coding for these sequences. ADNA coding for each CDR sequence may be determined on the basis of theamino acid sequence of the CDR and synthesized together with any desiredantibody variable region framework and constant region DNA sequencesusing oligonucleotide synthesis techniques, site-directed mutagenesisand polymerase chain reaction (PCR) techniques as appropriate. DNAcoding for variable region frameworks and constant regions is widelyavailable to those skilled in the art from genetic sequences databasessuch as GenBank®. Each of the above-mentioned CDRs will be typicallylocated in a variable region framework at positions 31-35 (CDR-H1),50-65 (CDR-H2) and 95-102 (CDR-H3) of the heavy chain and positions24-34 (CDR-L1), 50-56 (CDR-L2) and 89-97 (CDR-L3) of the light chainaccording to the Kabat numbering system (Kabat et al., 1987 in Sequencesof Proteins of Immunological Interest, U.S. Department of Health andHuman Services, NIH, USA).

Once synthesized, the DNA encoding an antibody of the invention orfragment thereof may be propagated and expressed according to any of avariety of well-known procedures for nucleic acid excision, ligation,transformation, and transfection using any number of known expressionvectors. Thus, in certain embodiments expression of an antibody fragmentmay be preferred in a prokaryotic host, such as Escherichia coli (see,e.g., Pluckthun et al., 1989 Methods Enzymol. 178:497-515). In certainother embodiments, expression of the antibody or a fragment thereof maybe preferred in a eukaryotic host cell, including yeast (e.g.,Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Pichiapastoris), animal cells (including mammalian cells) or plant cells.Examples of suitable animal cells include, but are not limited to,myeloma (such as a mouse NSO line), COS, CHO, or hybridoma cells.Examples of plant cells include tobacco, corn, soybean, and rice cells.

One or more replicable expression vectors containing DNA encoding anantibody variable and/or constant region may be prepared and used totransform an appropriate cell line, for example, a non-producing myelomacell line, such as a mouse NSO line or a bacteria, such as E. coli, inwhich production of the antibody will occur. In order to obtainefficient transcription and translation, the DNA sequence in each vectorshould include appropriate regulatory sequences, particularly a promoterand leader sequence operatively linked to the variable domain sequence.Particular methods for producing antibodies in this way are generallywell-known and routinely used. For example, basic molecular biologyprocedures are described by Maniatis et al. (Molecular Cloning, ALaboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, New York,1989; see also Maniatis et al, 3rd ed., Cold Spring Harbor Laboratory,New York, (2001)). DNA sequencing can be performed as described inSanger et al. (PNAS 74:5463, (1977)) and the Amersham International plcsequencing handbook, and site directed mutagenesis can be carried outaccording to methods known in the art (Kramer et al., Nucleic Acids Res.12:9441, (1984); Kunkel Proc. Natl. Acad. Sci. USA 82:488-92 (1985);Kunkel et al., Methods in Enzymol. 154:367-82 (1987); the AnglianBiotechnology Ltd handbook). Additionally, numerous publicationsdescribe techniques suitable for the preparation of antibodies bymanipulation of DNA, creation of expression vectors, and transformationand culture of appropriate cells (Mountain A and Adair, J R inBiotechnology and Genetic Engineering Reviews (ed. Tombs, M P, 10,Chapter 1, 1992, Intercept, Andover, UK); “Current Protocols inMolecular Biology”, 1999, F. M. Ausubel (ed.), Wiley Interscience, NewYork).

Where it is desired to improve the affinity of antibodies according tothe invention containing one or more of the above-mentioned CDRs can beobtained by a number of affinity maturation protocols includingmaintaining the CDRs (Yang et al., J. Mol. Biol., 254, 392-403, 1995),chain shuffling (Marks et al., Bio/Technology, 10, 779-783, 1992), useof mutation strains of E. coli. (Low et al., J. Mol. Biol., 250,350-368, 1996), DNA shuffling (Patten et al., Curr. Opin. Biotechnol.,8, 724-733, 1997), phage display (Thompson et al., J. Mol. Biol., 256,7-88, 1996) and sexual PCR (Crameri, et al., Nature, 391, 288-291,1998). All of these methods of affinity maturation are discussed byVaughan et al. (Nature Biotechnology, 16, 535-539, 1998).

Other antibodies according to the invention may be obtained byconventional immunization and cell fusion procedures as described hereinand known in the art. Monoclonal antibodies of the invention may begenerated using a variety of known techniques. In general, monoclonalantibodies that bind to specific antigens may be obtained by methodsknown to those skilled in the art (see, for example, Kohler et al.,Nature 256:495, 1975; Coligan et al. (eds.), Current Protocols inImmunology, 1:2.5.12.6.7 (John Wiley & Sons 1991); U.S. Pat. Nos. RE32,011, 4,902,614, 4,543,439, and 4,411,993; Monoclonal Antibodies,Hybridomas: A New Dimension in Biological Analyses, Plenum Press,Kennett, McKearn, and Bechtol (eds.) (1980); and Antibodies: ALaboratory Manual, Harlow and Lane (eds.), Cold Spring Harbor LaboratoryPress (1988); Picksley et al., “Production of monoclonal antibodiesagainst proteins expressed in E. coli,” in DNA Cloning 2: ExpressionSystems, 2nd Edition, Glover et al. (eds.), page 93 (Oxford UniversityPress 1995)). Antibody fragments may be derived therefrom using anysuitable standard technique such as proteolytic digestion, oroptionally, by proteolytic digestion (for example, using papain orpepsin) followed by mild reduction of disulfide bonds and alkylation.Alternatively, such fragments may also be generated by recombinantgenetic engineering techniques as described herein.

Monoclonal antibodies can be obtained by injecting an animal, forexample, a rat, hamster, a rabbit, or preferably a mouse, including forexample a transgenic or a knock-out, as known in the art, with animmunogen comprising human sclerostin of SEQ ID NO:1, or a fragmentthereof, according to methods known in the art and described herein. Thepresence of specific antibody production may be monitored after theinitial injection and/or after a booster injection by obtaining a serumsample and detecting the presence of an antibody that binds to humansclerostin or peptide using any one of several immunodetection methodsknown in the art and described herein. From animals producing thedesired antibodies, lymphoid cells, most commonly cells from the spleenor lymph node, are removed to obtain B-lymphocytes. The B lymphocytesare then fused with a drug-sensitized myeloma cell fusion partner,preferably one that is syngeneic with the immunized animal and thatoptionally has other desirable properties (e.g., inability to expressendogenous Ig gene products, e.g., P3X63—Ag 8.653 (ATCC No. CRL 1580);NSO, SP20) to produce hybridomas, which are immortal eukaryotic celllines. The lymphoid (e.g., spleen) cells and the myeloma cells may becombined for a few minutes with a membrane fusion-promoting agent, suchas polyethylene glycol or a nonionic detergent, and then plated at lowdensity on a selective medium that supports the growth of hybridomacells but not unfused myeloma cells. A preferred selection media is HAT(hypoxanthine, aminopterin, thymidine). After a sufficient time, usuallyabout one to two weeks, colonies of cells are observed. Single coloniesare isolated, and antibodies produced by the cells may be tested forbinding activity to human sclerostin, using any one of a variety ofimmunoassays known in the art and described herein. The hybridomas arecloned (e.g., by limited dilution cloning or by soft agar plaqueisolation) and positive clones that produce an antibody specific tosclerostin are selected and cultured. The monoclonal antibodies from thehybridoma cultures may be isolated from the supernatants of hybridomacultures. An alternative method for production of a murine monoclonalantibody is to inject the hybridoma cells into the peritoneal cavity ofa syngeneic mouse, for example, a mouse that has been treated (e.g.,pristane-primed) to promote formation of ascites fluid containing themonoclonal antibody. Monoclonal antibodies can be isolated and purifiedby a variety of well-established techniques. Such isolation techniquesinclude affinity chromatography with Protein-A Sepharose, size-exclusionchromatography, and ion-exchange chromatography (see, for example,Coligan at pages 2.7.1-2.7.12 and pages 2.9.1-2.9.3; Baines et al.,“Purification of Immunoglobulin G (IgG),” in Methods in MolecularBiology, Vol. 10, pages 79-104 (The Humana Press, Inc. 1992)).Monoclonal antibodies may be purified by affinity chromatography usingan appropriate ligand selected based on particular properties of theantibody (e.g., heavy or light chain isotype, binding specificity,etc.). Examples of a suitable ligand, immobilized on a solid support,include Protein A, Protein G, an anticonstant region (light chain orheavy chain) antibody, an anti-idiotype antibody, and a TGF-beta bindingprotein, or fragment or variant thereof.

An antibody of the present invention may also be a human monoclonalantibody. Human monoclonal antibodies may be generated by any number oftechniques with which those having ordinary skill in the art will befamiliar. Such methods include, but are not limited to, Epstein BarrVirus (EBV) transformation of human peripheral blood cells (e.g.,containing B lymphocytes), in vitro immunization of human B cells,fusion of spleen cells from immunized transgenic mice carrying insertedhuman immunoglobulin genes, isolation from human immunoglobulin V regionphage libraries, or other procedures as known in the art and based onthe disclosure herein. For example, human monoclonal antibodies may beobtained from transgenic mice that have been engineered to producespecific human antibodies in response to antigenic challenge. Methodsfor obtaining human antibodies from transgenic mice are described, forexample, by Green et al., Nature Genet. 7:13, 1994; Lonberg et al.,Nature 368:856, 1994; Taylor et al., Int. Immun. 6:579, 1994; U.S. Pat.No. 5,877,397; Bruggemann et al., 1997 Curr. Opin. Biotechnol. 8:455-58;Jakobovits et al., 1995 Ann. N. Y. Acad. Sci. 764:525-35. In thistechnique, elements of the human heavy and light chain locus areintroduced into strains of mice derived from embryonic stem cell linesthat contain targeted disruptions of the endogenous heavy chain andlight chain loci (see also Bruggemann et al., Curr. Opin. Biotechnol.8:455-58 (1997)). For example, human immunoglobulin transgenes may bemini-gene constructs, or transloci on yeast artificial chromosomes,which undergo B cell-specific DNA rearrangement and hypermutation in themouse lymphoid tissue. Human monoclonal antibodies may be obtained byimmunizing the transgenic mice, which may then produce human antibodiesspecific for sclerostin. Lymphoid cells of the immunized transgenic micecan be used to produce human antibody-secreting hybridomas according tothe methods described herein. Polyclonal sera containing humanantibodies may also be obtained from the blood of the immunized animals.

Another method for generating human antibodies of the invention includesimmortalizing human peripheral blood cells by EBV transformation. See,e.g., U.S. Pat. No. 4,464,456. Such an immortalized B cell line (orlymphoblastoid cell line) producing a monoclonal antibody thatspecifically binds to sclerostin can be identified by immunodetectionmethods as provided herein, for example, an ELISA, and then isolated bystandard cloning techniques. The stability of the lymphoblastoid cellline producing an anti-sclerostin antibody may be improved by fusing thetransformed cell line with a murine myeloma to produce a mouse-humanhybrid cell line according to methods known in the art (see, e.g.,Glasky et al., Hybridoma 8:377-89 (1989)). Still another method togenerate human monoclonal antibodies is in vitro immunization, whichincludes priming human splenic B cells with human sclerostin, followedby fusion of primed B cells with a heterohybrid fusion partner. See,e.g., Boerner et al., 1991. J. Immunol. 147:86-95.

In certain embodiments, a B cell that is producing an anti-humansclerostin antibody is selected and the light chain and heavy chainvariable regions are cloned from the B cell according to molecularbiology techniques known in the art (WO 92/02551; U.S. Pat. No.5,627,052; Babcook et al., Proc. Natl. Acad. Sci. USA 93:7843-48 (1996))and described herein. B cells from an immunized animal may be isolatedfrom the spleen, lymph node, or peripheral blood sample by selecting acell that is producing an antibody that specifically binds tosclerostin. B cells may also be isolated from humans, for example, froma peripheral blood sample. Methods for detecting single B cells that areproducing an antibody with the desired specificity are well known in theart, for example, by plaque formation, fluorescence-activated cellsorting, in vitro stimulation followed by detection of specificantibody, and the like. Methods for selection of specificantibody-producing B cells include, for example, preparing a single cellsuspension of B cells in soft agar that contains human sclerostin.Binding of the specific antibody produced by the B cell to the antigenresults in the formation of a complex, which may be visible as animmunoprecipitate. After the B cells producing the desired antibody areselected, the specific antibody genes may be cloned by isolating andamplifying DNA or mRNA according to methods known in the art anddescribed herein.

An additional method for obtaining antibodies of the invention is byphage display. See, e.g., Winter et al., 1994 Annu. Rev. Immunol.12:433-55; Burton et al., 1994 Adv. Immunol. 57:191-280. Human or murineimmunoglobulin variable region gene combinatorial libraries may becreated in phage vectors that can be screened to select Ig fragments(Fab, Fv, sFv, or multimers thereof) that bind specifically to TGF-betabinding protein or variant or fragment thereof. See, e.g., U.S. Pat. No.5,223,409; Huse et al., 1989 Science 246:1275-81; Sastry et al., Proc.Natl. Acad. Sci. USA 86:5728-32 (1989); Alting-Mees et al., Strategiesin Molecular Biology 3:1-9 (1990); Kang et al., 1991 Proc. Natl. Acad.Sci. USA 88:4363-66; Hoogenboom et al., 1992 J. Molec. Biol.227:381-388; Schlebusch et al., 1997 Hybridoma 16:47-52 and referencescited therein. For example, a library containing a plurality ofpolynucleotide sequences encoding Ig variable region fragments may beinserted into the genome of a filamentous bacteriophage, such as M13 ora variant thereof, in frame with the sequence encoding a phage coatprotein. A fusion protein may be a fusion of the coat protein with thelight chain variable region domain and/or with the heavy chain variableregion domain. According to certain embodiments, immunoglobulin Fabfragments may also be displayed on a phage particle (see, e.g., U.S.Pat. No. 5,698,426).

Heavy and light chain immunoglobulin cDNA expression libraries may alsobe prepared in lambda phage, for example, using λImmunoZap™(H) andλImmunoZap™(L) vectors (Stratagene, La Jolla, Calif.). Briefly, mRNA isisolated from a B cell population, and used to create heavy and lightchain immunoglobulin cDNA expression libraries in the λImmunoZap(H) andλImmunoZap(L) vectors. These vectors may be screened individually orco-expressed to form Fab fragments or antibodies (see Huse et al.,supra; see also Sastry et al., supra). Positive plaques may subsequentlybe converted to a non-lytic plasmid that allows high level expression ofmonoclonal antibody fragments from E. coli.

In one embodiment, in a hybridoma the variable regions of a geneexpressing a monoclonal antibody of interest are amplified usingnucleotide primers. These primers may be synthesized by one of ordinaryskill in the art, or may be purchased from commercially availablesources. (See, e.g., Stratagene (La Jolla, Calif.), which sells primersfor mouse and human variable regions including, among others, primersfor V_(Ha), V_(Hb), V_(Hc), V_(Hd), C_(Hl), V_(L) and C_(L) regions.)These primers may be used to amplify heavy or light chain variableregions, which may then be inserted into vectors such as ImmunoZAP™H orImmunoZAP™L (Stratagene), respectively. These vectors may then beintroduced into E. coli, yeast, or mammalian-based systems forexpression. Large amounts of a single-chain protein containing a fusionof the V_(H) and V_(L) domains may be produced using these methods (seeBird et al., Science 242:423-426, 1988).

Once cells producing antibodies according to the invention have beenobtained using any of the above-described immunization and othertechniques, the specific antibody genes may be cloned by isolating andamplifying DNA or mRNA therefrom according to standard procedures asdescribed herein. The antibodies produced therefrom may be sequenced andthe CDRs identified and the DNA coding for the CDRs may be manipulatedas described previously to generate other antibodies according to theinvention.

Preferably the binding agents specifically bind to sclerostin. As withall binding agents and binding assays, one of skill in this artrecognizes that the various moieties to which a binding agent should notdetectably bind in order to be therapeutically effective and suitablewould be exhaustive and impractical to list. Therefore, for a bindingagent disclosed herein, the term “specifically binds” refers to theability of a binding agent to bind to sclerostin, preferably humansclerostin, with greater affinity than it binds to an unrelated controlprotein. Preferably the control protein is hen egg white lysozyme.Preferably the binding agents bind to sclerostin with an affinity thatis at least, 50, 100, 250, 500, 1000, or 10,000 times greater than theaffinity for a control protein. A binding agent may have a bindingaffinity for human sclerostin of less than or equal to 1×10⁻⁷M, lessthan or equal to 1×10⁻⁸M, less than or equal to 1×10⁻⁹M, less than orequal to 1×10⁻¹⁰ M, less than or equal to 1×10⁻¹¹M, or less than orequal to 1×10⁻¹² M.

Affinity may be determined by an affinity ELISA assay. In certainembodiments, affinity may be determined by a BIAcore assay. In certainembodiments, affinity may be determined by a kinetic method. In certainembodiments, affinity may be determined by an equilibrium/solutionmethod. Such methods are described in further detail herein or known inthe art.

Sclerostin binding agents of the present invention preferably modulatesclerostin function in the cell-based assay described herein and/or thein vivo assay described herein and/or bind to one or more of theepitopes described herein and/or cross-block the binding of one of theantibodies described in this application and/or are cross-blocked frombinding sclerostin by one of the antibodies described in thisapplication. Accordingly such binding agents can be identified using theassays described herein.

In certain embodiments, binding agents are generated by firstidentifying antibodies that bind to one more of the epitopes providedherein and/or neutralize in the cell-based and/or in vivo assaysdescribed herein and/or cross-block the antibodies described in thisapplication and/or are cross-blocked from binding sclerostin by one ofthe antibodies described in this application. The CDR regions from theseantibodies are then used to insert into appropriate biocompatibleframeworks to generate sclerostin binding agents. The non-CDR portion ofthe binding agent may be composed of amino acids, or may be anon-protein molecule. The assays described herein allow thecharacterization of binding agents. Preferably the binding agents of thepresent invention are antibodies as defined herein.

It will be understood by one skilled in the art that some proteins, suchas antibodies, may undergo a variety of posttranslational modifications.The type and extent of these modifications often depends on the hostcell line used to express the protein as well as the culture conditions.Such modifications may include variations in glycosylation, methionineoxidation, diketopiperizine formation, aspartate isomerization andasparagine deamidation. A frequent modification is the loss of acarboxy-terminal basic residue (such as lysine or arginine) due to theaction of carboxypeptidases (as described in Harris, R J. Journal ofChromatography 705:129-134, 1995).

Antibodies referred to as Ab-A, Ab-B, Ab-C, Ab-D and Ab-1 are describedbelow. “HC” refers to the heavy chain and “LC” refers to the lightchain. For some antibodies below, the CDRs are box shaded and theconstant (C) regions are shown in bold italics.

Ab-D

Antibody D (also referred to herein as Ab-D and Mab-D) is a mouseantibody which exhibits high affinity binding to sclerostin. The BIAcorebinding pattern of Ab-D is shown in FIG. 18.

The amino acid sequence of the mature form (signal peptide removed) ofAb-D light chain:

(SEQ ID NO: 7)

Nucleic acid sequence encoding the mature form (signal peptide removed)of Ab-D LC is as follows:

(SEQ ID NO: 8) 1 GATGTCCAGA TGATTCAGTC TCCATCCTCC CTGTCTGCAT CTTTGGGAGA51 CATAGTCACC ATGACTTGCC AGGCAAGTCA GGGCACTAGC ATTAATTTAA 101 ACTGGTTTCAGCAAAAACCA GGGAAGGCTC CTAAGCTCCT GATCTATGGT 151 TCAAGCAACT TGGAAGATGGGGTCCCATCA AGGTTCAGTG GCAGTAGATA 201 TGGGACAGAT TTCACTCTCA CCATCAGCAGCCTGGAGGAT GAAGATCTGG 251 CAACTTATTT CTGTCTACAA CATAGTTATC TCCCGTACACGTTCGGAGGG 301 GGGACCAAGC TGGAAATAAA ACGGGCTGAT GCTGCACCAA CTGTATCCAT351 CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC TCAGTCGTGT 401GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA GTGGAAGATT 451 GATGGCAGTGAACGACAAAA TGGCGTCCTG AACAGTTGGA CTGATCAGGA 501 CAGCAAAGAC AGCACCTACAGCATGAGCAG CACCCTCACG TTGACCAAGG 551 ACGAGTATGA ACGACATAAC AGCTATACCTGTGAGGCCAC TCACAAGACA 601 TCAACTTCAC CCATTGTCAA GAGCTTCAAC AGGAATGAGTGTTAG

The amino acid sequence of Ab-D LC including signal peptide is asfollows:

(SEQ ID NO: 9) 1 MNTRAPAEFL GFLLLWFLGA RCDVQMIQSP SSLSASLGDI VTMTCQASQG51 TSINLNWFQQ KPGKAPKLLI YGSSNLEDGV PSRFSGSRYG TDFTLTISSL 101 EDEDLATYFCLQHSYLPYTF GGGTKLEIKR ADAAPTVSIF PPSSEQLTSG 151 GASVVCFLNN FYPKDINVKWKIDGSERQNG VLNSWTDQDS KDSTYSMSST 201 LTLTKDEYER HNSYTCEATH KTSTSPIVKSFNRNEC

Nucleic acid sequence of Ab-D LC including signal peptide encodingsequence:

(SEQ ID NO: 10) 1 ATGAACACGA GGGCCCCTGC TGAGTTCCTT GGGTTCCTGT TGCTCTGGTT51 TTTAGGTGCC AGATGTGATG TCCAGATGAT TCAGTCTCCA TCCTCCCTGT 101 CTGCATCTTTGGGAGACATA GTCACCATGA CTTGCCAGGC AAGTCAGGGC 151 ACTAGCATTA ATTTAAACTGGTTTCAGCAA AAACCAGGGA AGGCTCCTAA 201 GCTCCTGATC TATGGTTCAA GCAACTTGGAAGATGGGGTC CCATCAAGGT 251 TCAGTGGCAG TAGATATGGG ACAGATTTCA CTCTCACCATCAGCAGCCTG 301 GAGGATGAAG ATCTGGCAAC TTATTTCTGT CTACAACATA GTTATCTCCC351 GTACACGTTC GGAGGGGGGA CCAAGCTGGA AATAAAACGG GCTGATGCTG 401CACCAACTGT ATCCATCTTC CCACCATCCA GTGAGCAGTT AACATCTGGA 451 GGTGCCTCAGTCGTGTGCTT CTTGAACAAC TTCTACCCCA AAGACATCAA 501 TGTCAAGTGG AAGATTGATGGCAGTGAACG ACAAAATGGC GTCCTGAACA 551 GTTGGACTGA TCAGGACAGC AAAGACAGCACCTACAGCAT GAGCAGCACC 601 CTCACGTTGA CCAAGGACGA GTATGAACGA CATAACAGCTATACCTGTGA 651 GGCCACTCAC AAGACATCAA CTTCACCCAT TGTCAAGAGC TTCAACAGGA701 ATGAGTGTTA G

The amino acid sequence of the mature form (signal peptide removed) ofAb-D HC heavy chain is as follows:

(SEQ ID NO: 11)

The nucleic acid sequence encoding the mature form (signal peptideremoved) of Ab-D HC is:

(SEQ ID NO: 12) 1 GAGGTCCAGC TGCAACAGTC TGGACCTGAA CTGGTGACGC CTGGGGCTTC51 AGTGAAGATA TCTTGTAAGG CTTCTGGATA CACATTCACT GACCACTACA 101 TGAGCTGGGTGAAGCAGAGT CATGGAAAAA GCCTTGAGTG GATTGGAGAT 151 ATTAATCCCT ATTCTGGTGAAACTACCTAC AACCAGAAGT TCAAGGGCAC 201 GGCCACATTG ACTGTAGACA AGTCTTCCAGTATAGCCTAC ATGGAGATCC 251 GCGGCCTGAC ATCTGAGGAC TCTGCAGTCT ATTACTGTGCAAGAGATGAT 301 TACGACGCCT CTCCGTTTGC TTACTGGGGC CAAGGGACTC TGGTCACTGT351 CTCTGCAGCC AAAACGACAC CCCCATCTGT CTATCCACTG GCCCCTGGAT 401CTGCTGCCCA AACTAACTCC ATGGTGACCC TGGGATGCCT GGTCAAGGGC 451 TATTTCCCTGAGCCAGTGAC AGTGACCTGG AACTCTGGAT CCCTGTCCAG 501 CGGTGTGCAC ACCTTCCCAGCTGTCCTGCA GTCTGACCTC TACACTCTGA 551 GCAGCTCAGT GACTGTCCCC TCCAGCACCTGGCCCAGCGA GACCGTCACC 601 TGCAACGTTG CCCACCCGGC CAGCAGCACC AAGGTGGACAAGAAAATTGT 651 GCCCAGGGAT TGTGGTTGTA AGCCTTGCAT ATGTACAGTC CCAGAAGTAT701 CATCTGTCTT CATCTTCCCC CCAAAGCCCA AGGATGTGCT CACCATTACT 751CTGACTCCTA AGGTCACGTG TGTTGTGGTA GACATCAGCA AGGATGATCC 801 CGAGGTCCAGTTCAGCTGGT TTGTAGATGA TGTGGAGGTG CACACAGCTC 851 AGACGCAACC CCGGGAGGAGCAGTTCAACA GCACTTTCCG CTCAGTCAGT 901 GAACTTCCCA TCATGCACCA GGACTGGCTCAATGGCAAGG AGTTCAAATG 951 CAGGGTCAAC AGTCCAGCTT TCCCTGCCCC CATCGAGAAAACCATCTCCA 1001 AAACCAAAGG CAGACCGAAG GCTCCACAGG TGTACACCAT TCCACCTCCC1051 AAGGAGCAGA TGGCCAAGGA TAAAGTCAGT CTGACCTGCA TGATAACAGA 1101CTTCTTCCCT GAAGACATTA CTGTGGAGTG GCAGTGGAAT GGGCAGCCAG 1151 CGGAGAACTACAAGAACACT CAGCCCATCA TGGACACAGA TGGCTCTTAC 1201 TTCATCTACA GCAAGCTCAATGTGCAGAAG AGCAACTGGG AGGCAGGAAA 1251 TACTTTCACC TGCTCTGTGT TACATGAGGGCCTGCACAAC CACCATACTG 1301 AGAAGAGCCT CTCCCACTCT CCTGGTAAAT GA

The amino acid sequence of Ab-D HC including signal peptide is:

(SEQ ID NO: 13) 1 MRCRWIFLFL LSGTAGVLSE VQLQQSGPEL VTPGASVKIS CKASGYTFTD51 HYMSWVKQSH GKSLEWIGDI NPYSGETTYN QKFKGTATLT VDKSSSIAYM 101 EIRGLTSEDSAVYYCARDDY DASPFAYWGQ GTLVTVSAAK TTPPSVYPLA 151 PGSAAQTNSM VTLGCLVKGYFPEPVTVTWN SGSLSSGVHT FPAVLQSDLY 201 TLSSSVTVPS STWPSETVTC NVAHPASSTKVDKKIVPRDC GCKPCICTVP 251 EVSSVFIFPP KPKDVLTITL TPKVTCVVVD ISKDDPEVQFSWFVDDVEVH 301 TAQTQPREEQ FNSTFRSVSE LPIMHQDWLN GKEFKCRVNS PAFPAPIEKT351 ISKTKGRPKA PQVYTIPPPK EQMAKDKVSL TCMITDFFPE DITVEWQWNG 401QPAENYKNTQ PIMDTDGSYF IYSKLNVQKS NWEAGNTFTC SVLHEGLHNH 451 HTEKSLSHSP GK

The nucleic acid sequence of Ab-D HC including signal peptide encodingsequence is:

(SEQ ID NO: 14) 1 ATGAGATGCA GGTGGATCTT TCTCTTTCTC CTGTCAGGAA CTGCAGGTGT51 CCTCTCTGAG GTCCAGCTGC AACAGTCTGG ACCTGAACTG GTGACGCCTG 101 GGGCTTCAGTGAAGATATCT TGTAAGGCTT CTGGATACAC ATTCACTGAC 151 CACTACATGA GCTGGGTGAAGCAGAGTCAT GGAAAAAGCC TTGAGTGGAT 201 TGGAGATATT AATCCCTATT CTGGTGAAACTACCTACAAC CAGAAGTTCA 251 AGGGCACGGC CACATTGACT GTAGACAAGT CTTCCAGTATAGCCTACATG 301 GAGATCCGCG GCCTGACATC TGAGGACTCT GCAGTCTATT ACTGTGCAAG351 AGATGATTAC GACGCCTCTC CGTTTGCTTA CTGGGGCCAA GGGACTCTGG 401TCACTGTCTC TGCAGCCAAA ACGACACCCC CATCTGTCTA TCCACTGGCC 451 CCTGGATCTGCTGCCCAAAC TAACTCCATG GTGACCCTGG GATGCCTGGT 501 CAAGGGCTAT TTCCCTGAGCCAGTGACAGT GACCTGGAAC TCTGGATCCC 551 TGTCCAGCGG TGTGCACACC TTCCCAGCTGTCCTGCAGTC TGACCTCTAC 601 ACTCTGAGCA GCTCAGTGAC TGTCCCCTCC AGCACCTGGCCCAGCGAGAC 651 CGTCACCTGC AACGTTGCCC ACCCGGCCAG CAGCACCAAG GTGGACAAGA701 AAATTGTGCC CAGGGATTGT GGTTGTAAGC CTTGCATATG TACAGTCCCA 751GAAGTATCAT CTGTCTTCAT CTTCCCCCCA AAGCCCAAGG ATGTGCTCAC 801 CATTACTCTGACTCCTAAGG TCACGTGTGT TGTGGTAGAC ATCAGCAAGG 851 ATGATCCCGA GGTCCAGTTCAGCTGGTTTG TAGATGATGT GGAGGTGCAC 901 ACAGCTCAGA CGCAACCCCG GGAGGAGCAGTTCAACAGCA CTTTCCGCTC 951 AGTCAGTGAA CTTCCCATCA TGCACCAGGA CTGGCTCAATGGCAAGGAGT 1001 TCAAATGCAG GGTCAACAGT CCAGCTTTCC CTGCCCCCAT CGAGAAAACC1051 ATCTCCAAAA CCAAAGGCAG ACCGAAGGCT CCACAGGTGT ACACCATTCC 1101ACCTCCCAAG GAGCAGATGG CCAAGGATAA AGTCAGTCTG ACCTGCATGA 1151 TAACAGACTTCTTCCCTGAA GACATTACTG TGGAGTGGCA GTGGAATGGG 1201 CAGCCAGCGG AGAACTACAAGAACACTCAG CCCATCATGG ACACAGATGG 1251 CTCTTACTTC ATCTACAGCA AGCTCAATGTGCAGAAGAGC AACTGGGAGG 1301 CAGGAAATAC TTTCACCTGC TCTGTGTTAC ATGAGGGCCTGCACAACCAC 1351 CATACTGAGA AGAGCCTCTC CCACTCTCCT GGTAAATGA

The CDR (complementarity determining region) sequences in the variableregion of the heavy chain of Ab-D are as follows:

DR-H1: (SEQ ID NO: 39) DHYMS CDR-H2: (SEQ ID NO: 40) DINPYSGETTYNQKFKGCDR-H3: (SEQ ID NO: 41) DDYDASPFAY

-   -   The light chain variable region CDR sequences of Ab-D are:

(SEQ ID NO: 42) CDR-L1: QASQGTSINLN (SEQ ID NO: 43) CDR-L2: GSSNLED (SEQID NO: 44) CDR-L3: LQHSYLPYT

Ab-C

Antibody C (also referred to herein as Ab-C and Mab-C) is a mouseantibody which exhibits high affinity binding to sclerostin. The BIAcorebinding pattern of Ab-C is shown in FIG. 17. The amino acid sequence ofthe mature form (signal peptide removed) of Ab-C Light Chain is asfollows:

(SEQ ID NO: 15)

The nucleic acid sequence encoding the mature form (signal peptideremoved) of Ab-C LC is:

(SEQ ID NO: 16) 1 GACATTGTGC TGACCCAATC TCCAGCTTCT TTGACTGTGT CTCTAGGCCT51 GAGGGCCACC ATCTCCTGCA AGGCCAGCCA AAGTGTTGAT TATGATGGTG 101 ATAGTTATATGAACTGGTAC CAGCAGAAAC CAGGACAGCC ACCCAAACTC 151 CTCATCTATG CTGCATCCAATCTAGAATCT GGGATCCCAG CCAGGTTTAG 201 TGGCAATGGG TCTGGGACAG ACTTCACCCTCAACATCCAT CCTGTGGAGG 251 AGGAGGATGC TGTAACCTAT TACTGTCAAC AAAGTAATGAGGATCCGTGG 301 ACGTTCGGTG GAGGCACCAA GCTGGAAATC AAACGGGCTG ATGCTGCACC351 AACTGTATCC ATCTTCCCAC CATCCAGTGA GCAGTTAACA TCTGGAGGTG 401CCTCAGTCGT GTGCTTCTTG AACAACTTCT ACCCCAAAGA CATCAATGTC 451 AAGTGGAAGATTGATGGCAG TGAACGACAA AATGGCGTCC TGAACAGTTG 501 GACTGATCAG GACAGCAAAGACAGCACCTA CAGCATGAGC AGCACCCTCA 551 CGTTGACCAA GGACGAGTAT GAACGACATAACAGCTATAC CTGTGAGGCC 601 ACTCACAAGA CATCAACTTC ACCCATTGTC AAGAGCTTCAACAGGAATGA 651 GTGTTAG

The amino acid sequence of Ab-C LC including signal peptide is:

(SEQ ID NO: 17) 1 METDTILLWV LLLWVPGSTG DIVLTQSPAS LTVSLGLRAT ISCKASQSVD51 YDGDSYMNWY QQKPGQPPKL LIYAASNLES GIPARFSGNG SGTDFTLNIH 101 PVEEEDAVTYYCQQSNEDPW TFGGGTKLEI KRADAAPTVS IFPPSSEQLT 151 SGGASVVCFL NNFYPKDINVKWKIDGSERQ NGVLNSWTDQ DSKDSTYSMS 201 STLTLTKDEY ERHNSYTCEA THKTSTSPIVKSFNRNEC

The nucleic acid sequence of Ab-C LC including signal peptide encodingsequence is:

(SEQ ID NO: 18) 1 ATGGAGACAG ACACAATCCT GCTATGGGTG CTGCTGCTCT GGGTTCCAGG51 CTCCACTGGT GACATTGTGC TGACCCAATC TCCAGCTTCT TTGACTGTGT 101 CTCTAGGCCTGAGGGCCACC ATCTCCTGCA AGGCCAGCCA AAGTGTTGAT 151 TATGATGGTG ATAGTTATATGAACTGGTAC CAGCAGAAAC CAGGACAGCC 201 ACCCAAACTC CTCATCTATG CTGCATCCAATCTAGAATCT GGGATCCCAG 251 CCAGGTTTAG TGGCAATGGG TCTGGGACAG ACTTCACCCTCAACATCCAT 301 CCTGTGGAGG AGGAGGATGC TGTAACCTAT TACTGTCAAC AAAGTAATGA351 GGATCCGTGG ACGTTCGGTG GAGGCACCAA GCTGGAAATC AAACGGGCTG 401ATGCTGCACC AACTGTATCC ATCTTCCCAC CATCCAGTGA GCAGTTAACA 451 TCTGGAGGTGCCTCAGTCGT GTGCTTCTTG AACAACTTCT ACCCCAAAGA 501 CATCAATGTC AAGTGGAAGATTGATGGCAG TGAACGACAA AATGGCGTCC 551 TGAACAGTTG GACTGATCAG GACAGCAAAGACAGCACCTA CAGCATGAGC 601 AGCACCCTCA CGTTGACCAA GGACGAGTAT GAACGACATAACAGCTATAC 651 CTGTGAGGCC ACTCACAAGA CATCAACTTC ACCCATTGTC AAGAGCTTCA701 ACAGGAATGA GTGTTAG

Ab-C Heavy Chain

The amino acid sequence of the mature form (signal peptide removed) ofAb-C HC is:

(SEQ ID NO: 19)

The nucleic acid sequence encoding the mature form (signal peptideremoved) of Ab-C HC is as follows:

(SEQ ID NO: 20) 1 GAGGTCCAGC TGCAACAATC TGGACCTGAG CTGGTGAAGC CTGGGACTTC51 AGTGAAGATG TCCTGTAAGG CTTCTGGATA CACATTCACT GACTGCTACA 101 TGAACTGGGTGAAGCAGAGC CATGGGAAGA GCCTTGAATG GATTGGAGAT 151 ATTAATCCTT TCAACGGTGGTACTACCTAC AACCAGAAGT TCAAGGGCAA 201 GGCCACATTG ACTGTAGACA AATCCTCCAGCACAGCCTAC ATGCAGCTCA 251 ACAGCCTGAC ATCTGACGAC TCTGCAGTCT ATTACTGTGCAAGATCCCAT 301 TATTACTTCG ATGGTAGAGT CCCTTGGGAT GCTATGGACT ACTGGGGTCA351 AGGAACCTCA GTCACCGTCT CCTCAGCCAA AACGACACCC CCATCTGTCT 401ATCCACTGGC CCCTGGATCT GCTGCCCAAA CTAACTCCAT GGTGACCCTG 451 GGATGCCTGGTCAAGGGCTA TTTCCCTGAG CCAGTGACAG TGACCTGGAA 501 CTCTGGATCC CTGTCCAGCGGTGTGCACAC CTTCCCAGCT GTCCTGCAGT 551 CTGACCTCTA CACTCTGAGC AGCTCAGTGACTGTCCCCTC CAGCACCTGG 601 CCCAGCGAGA CCGTCACCTG CAACGTTGCC CACCCGGCCAGCAGCACCAA 651 GGTGGACAAG AAAATTGTGC CCAGGGATTG TGGTTGTAAG CCTTGCATAT701 GTACAGTCCC AGAAGTATCA TCTGTCTTCA TCTTCCCCCC AAAGCCCAAG 751GATGTGCTCA CCATTACTCT GACTCCTAAG GTCACGTGTG TTGTGGTAGA 801 CATCAGCAAGGATGATCCCG AGGTCCAGTT CAGCTGGTTT GTAGATGATG 851 TGGAGGTGCA CACAGCTCAGACGCAACCCC GGGAGGAGCA GTTCAACAGC 901 ACTTTCCGCT CAGTCAGTGA ACTTCCCATCATGCACCAGG ACTGGCTCAA 951 TGGCAAGGAG TTCAAATGCA GGGTCAACAG TGCAGCTTTCCCTGCCCCCA 1001 TCGAGAAAAC CATCTCCAAA ACCAAAGGCA GACCGAAGGC TCCACAGGTG1051 TACACCATTC CACCTCCCAA GGAGCAGATG GCCAAGGATA AAGTCAGTCT 1101GACCTGCATG ATAACAGACT TCTTCCCTGA AGACATTACT GTGGAGTGGC 1151 AGTGGAATGGGCAGCCAGCG GAGAACTACA AGAACACTCA GCCCATCATG 1201 GACACAGATG GCTCTTACTTCATCTACAGC AAGCTCAATG TGCAGAAGAG 1251 CAACTGGGAG GCAGGAAATA CTTTCACCTGCTCTGTGTTA CATGAGGGCC 1301 TGCACAACCA CCATACTGAG AAGAGCCTCT CCCACTCTCCTGGTAAATGA

The amino acid sequence of Ab-C HC including signal peptide is:

(SEQ ID NO: 21) 1 MGWNWIFLFL LSGTAGVYSE VQLQQSGPEL VKPGTSVKMS CKASGYTFTD51 CYMNWVKQSH GKSLEWIGDI NPFNGGTTYN QKFKGKATLT VDKSSSTAYM 101 QLNSLTSDDSAVYYCARSHY YFDGRVPWDA MDYWGQGTSV TVSSAKTTPP 151 SVYPLAPGSA AQTNSMVTLGCLVKGYFPEP VTVTWNSGSL SSGVHTFPAV 201 LQSDLYTLSS SVTVPSSTWP SETVTCNVAHPASSTKVDKK IVPRDCGCKP 251 CICTVPEVSS VFIFPPKPKD VLTITLTPKV TCVVVDISKDDPEVQFSWFV 301 DDVEVHTAQT QPREEQFNST FRSVSELPIM HQDWLNGKEF KCRVNSAAFP351 APIEKTISKT KGRPKAPQVY TIPPPKEQMA KDKVSLTCMI TDFFPEDITV 401EWQWNGQPAE NYKNTQPIMD TDGSYFIYSK LNVQKSNWEA GNTFTCSVLH 451 EGLHNHHTEKSLSHSPGK

The nucleic acid sequence of Ab-C HC including signal peptide encodingsequence is:

(SEQ ID NO: 22) 1 ATGGGATGGA ACTGGATCTT TCTCTTCCTC TTGTCAGGAA CTGCAGGTGT51 CTACTCTGAG GTCCAGCTGC AACAATCTGG ACCTGAGCTG GTGAAGCCTG 101 GGACTTCAGTGAAGATGTCC TGTAAGGCTT CTGGATACAC ATTCACTGAC 151 TGCTACATGA ACTGGGTGAAGCAGAGCCAT GGGAAGAGCC TTGAATGGAT 201 TGGAGATATT AATCCTTTCA ACGGTGGTACTACCTACAAC CAGAAGTTCA 251 AGGGCAAGGC CACATTGACT GTAGACAAAT CCTCCAGCACAGCCTACATG 301 CAGCTCAACA GCCTGACATC TGACGACTCT GCAGTCTATT ACTGTGCAAG351 ATCCCATTAT TACTTCGATG GTAGAGTCCC TTGGGATGCT ATGGACTACT 401GGGGTCAAGG AACCTCAGTC ACCGTCTCCT CAGCCAAAAC GACACCCCCA 451 TCTGTCTATCCACTGGCCCC TGGATCTGCT GCCCAAACTA ACTCCATGGT 501 GACCCTGGGA TGCCTGGTCAAGGGCTATTT CCCTGAGCCA GTGACAGTGA 551 CCTGGAACTC TGGATCCCTG TCCAGCGGTGTGCACACCTT CCCAGCTGTC 601 CTGCAGTCTG ACCTCTACAC TCTGAGCAGC TCAGTGACTGTCCCCTCCAG 651 CACCTGGCCC AGCGAGACCG TCACCTGCAA CGTTGCCCAC CCGGCCAGCA701 GCACCAAGGT GGACAAGAAA ATTGTGCCCA GGGATTGTGG TTGTAAGCCT 751TGCATATGTA CAGTCCCAGA AGTATCATCT GTCTTCATCT TCCCCCCAAA 801 GCCCAAGGATGTGCTCACCA TTACTCTGAC TCCTAAGGTC ACGTGTGTTG 851 TGGTAGACAT CAGCAAGGATGATCCCGAGG TCCAGTTCAG CTGGTTTGTA 901 GATGATGTGG AGGTGCACAC AGCTCAGACGCAACCCCGGG AGGAGCAGTT 951 CAACAGCACT TTCCGCTCAG TCAGTGAACT TCCCATCATGCACCAGGACT 1001 GGCTCAATGG CAAGGAGTTC AAATGCAGGG TCAACAGTGC AGCTTTCCCT1051 GCCCCCATCG AGAAAACCAT CTCCAAAACC AAAGGCAGAC CGAAGGCTCC 1101ACAGGTGTAC ACCATTCCAC CTCCCAAGGA GCAGATGGCC AAGGATAAAG 1151 TCAGTCTGACCTGCATGATA ACAGACTTCT TCCCTGAAGA CATTACTGTG 1201 GAGTGGCAGT GGAATGGGCAGCCAGCGGAG AACTACAAGA ACACTCAGCC 1251 CATCATGGAC ACAGATGGCT CTTACTTCATCTACAGCAAG CTCAATGTGC 1301 AGAAGAGCAA CTGGGAGGCA GGAAATACTT TCACCTGCTCTGTGTTACAT 1351 GAGGGCCTGC ACAACCACCA TACTGAGAAG AGCCTCTCCC ACTCTCCTGG1401 TAAATGA

The CDR (complementarity determining region) sequences in the variableregion of the heavy chain of Ab-C are as follows:

(SEQ ID NO: 45) CDR-H1: DCYMN (SEQ ID NO: 46) CDR-H2: DINPFNGGTTYNQKFKG(SEQ ID NO: 47) CDR-H3: SHYYFDGRVPWDAMDY

-   -   The light chain variable region CDR sequences of Ab-C are:

(SEQ ID NO: 48) CDR-L1: KASQSVDYDGDSYMN (SEQ ID NO: 49) CDR-L2: AASNLES(SEQ ID NO: 50) CDR-L3: QQSNEDPWT

Ab-A

Antibody A (also referred to herein as Ab-A and Mab-A) is a rabbit-mousechimeric antibody which exhibits high affinity binding to sclerostin.The BIAcore binding pattern of Ab-A is shown in FIG. 15.

Ab-A Light Chain

The amino acid sequence of the mature form (signal peptide removed) ofAb-A LC:

(SEQ ID NO: 23)

The nucleic acid sequence encoding the mature form (signal peptideremoved) of Ab-A LC:

(SEQ ID NO: 24) 1 GCGCAAGTGC TGACCCAGAC TCCAGCCTCC GTGTCTGCAG CTGTGGGAGG51 CACAGTCACC ATCAATTGCC AGTCCAGTCA GAGTGTTTAT GATAACAACT 101 GGTTAGCCTGGTTTCAGCAG AAACCAGGGC AGCCTCCCAA GCTCCTGATT 151 TATGATGCAT CCGATCTGGCATCTGGGGTC CCATCGCGGT TCAGTGGCAG 201 TGGATCTGGG ACACAGTTCA CTCTCACCATCAGCGGCGTG CAGTGTGCCG 251 ATGCTGCCAC TTACTACTGT CAAGGCGCTT ATAATGATGTTATTTATGCT 301 TTCGGCGGAG GGACCGAGGT GGTGGTCAAA CGTACGGATG CTGCACCAAC351 TGTATCCATC TTCCCACCAT CCAGTGAGCA GTTAACATCT GGAGGTGCCT 401CAGTCGTGTG CTTCTTGAAC AACTTCTACC CCAAAGACAT CAATGTCAAG 451 TGGAAGATTGATGGCAGTGA ACGACAAAAT GGCGTCCTGA ACAGTTGGAC 501 TGATCAGGAC AGCAAAGACAGCACCTACAG CATGAGCAGC ACCCTCACGT 551 TGACCAAGGA CGAGTATGAA CGACATAACAGCTATACCTG TGAGGCCACT 601 CACAAGACAT CAACTTCACC CATTGTCAAG AGCTTCAACAGGAATGAGTG 651 TTAG

The amino acid sequence of Ab-A LC including signal peptide is:

(SEQ ID NO: 25) 1 MDTRAPTQLL GLLLLWLPGA TFAQVLTQTP ASVSAAVGGT VTINCQSSQS51 VYDNNWLAWF QQKPGQPPKL LIYDASDLAS GVPSRFSGSG SGTQFTLTIS 101 GVQCADAATYYCQGAYNDVI YAFGGGTEVV VKRTDAAPTV SIFPPSSEQL 151 TSGGASVVCF LNNFYPKDINVKWKIDGSER QNGVLNSWTD QDSKDSTYSM 201 SSTLTLTKDE YERHNSYTCE ATHKTSTSPIVKSFNRNEC

The nucleic acid sequence of Ab-A LC including signal peptide encodingsequence is:

(SEQ ID NO: 26) 1 ATGGACACGA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC TGCTCTGGCT51 CCCAGGTGCC ACATTTGCGC AAGTGCTGAC CCAGACTCCA GCCTCCGTGT 101 CTGCAGCTGTGGGAGGCACA GTCACCATCA ATTGCCAGTC CAGTCAGAGT 151 GTTTATGATA ACAACTGGTTAGCCTGGTTT CAGCAGAAAC CAGGGCAGCC 201 TCCCAAGCTC CTGATTTATG ATGCATCCGATCTGGCATCT GGGGTCCCAT 251 CGCGGTTCAG TGGCAGTGGA TCTGGGACAC AGTTCACTCTCACCATCAGC 301 GGCGTGCAGT GTGCCGATGC TGCCACTTAC TACTGTCAAG GCGCTTATAA351 TGATGTTATT TATGCTTTCG GCGGAGGGAC CGAGGTGGTG GTCAAACGTA 401CGGATGCTGC ACCAACTGTA TCCATCTTCC CACCATCCAG TGAGCAGTTA 451 ACATCTGGAGGTGCCTCAGT CGTGTGCTTC TTGAACAACT TCTACCCCAA 501 AGACATCAAT GTCAAGTGGAAGATTGATGG CAGTGAACGA CAAAATGGCG 551 TCCTGAACAG TTGGACTGAT CAGGACAGCAAAGACAGCAC CTACAGCATG 601 AGCAGCACCC TCACGTTGAC CAAGGACGAG TATGAACGACATAACAGCTA 651 TACCTGTGAG GCCACTCACA AGACATCAAC TTCACCCATT GTCAAGAGCT701 TCAACAGGAA TGAGTGTTAG

The amino acid sequence of the mature form (signal peptide removed) ofAb-A HC is:

(SEQ ID NO: 27)

The nucleic acid sequence encoding the mature form (signal peptideremoved) of Ab-A HC:

(SEQ ID NO: 28) 1 CAGTCGCTGG AGGAGTCCGG GGGTCGCCTG GTCACGCCTG GGACACCCCT51 GACACTCACC TGCACAGCCT CTGGATTCTC CCTCAGTAGT TATTGGATGA 101 ACTGGGTCCGCCAGGCTCCA GGGGAGGGGC TGGAATGGAT CGGAACCATT 151 GATTCTGGTG GTAGGACGGACTACGCGAGC TGGGCAAAAG GCCGATTCAC 201 CATCTCCAGA ACCTCGACTA CGATGGATCTGAAAATGACC AGTCTGACGA 251 CCGGGGACAC GGCCCGTTAT TTCTGTGCCA GAAATTGGAACTTGTGGGGC 301 CAAGGCACCC TCGTCACCGT CTCGAGCGCT TCTACAAAGG GCCCATCTGT351 CTATCCACTG GCCCCTGGAT CTGCTGCCCA AACTAACTCC ATGGTGACCC 401TGGGATGCCT GGTCAAGGGC TATTTCCCTG AGCCAGTGAC AGTGACCTGG 451 AACTCTGGATCCCTGTCCAG CGGTGTGCAC ACCTTCCCAG CTGTCCTGCA 501 GTCTGACCTC TACACTCTGAGCAGCTCAGT GACTGTCCCC TCCAGCACCT 551 GGCCCAGCGA GACCGTCACC TGCAACGTTGCCCACCCGGC CAGCAGCACC 601 AAGGTGGACA AGAAAATTGT GCCCAGGGAT TGTGGTTGTAAGCCTTGCAT 651 ATGTACAGTC CCAGAAGTAT CATCTGTCTT CATCTTCCCC CCAAAGCCCA701 AGGATGTGCT CACCATTACT CTGACTCCTA AGGTCACGTG TGTTGTGGTA 751GACATCAGCA AGGATGATCC CGAGGTCCAG TTCAGCTGGT TTGTAGATGA 801 TGTGGAGGTGCACACAGCTC AGACGCAACC CCGGGAGGAG CAGTTCAACA 851 GCACTTTCCG CTCAGTCAGTGAACTTCCCA TCATGCACCA GGACTGGCTC 901 AATGGCAAGG AGTTCAAATG CAGGGTCAACAGTGCAGCTT TCCCTGCCCC 951 CATCGAGAAA ACCATCTCCA AAACCAAAGG CAGACCGAAGGCTCCACAGG 1001 TGTACACCAT TCCACCTCCC AAGGAGCAGA TGGCCAAGGA TAAAGTCAGT1051 CTGACCTGCA TGATAACAGA CTTCTTCCCT GAAGACATTA CTGTGGAGTG 1101GCAGTGGAAT GGGCAGCCAG CGGAGAACTA CAAGAACACT CAGCCCATCA 1151 TGGACACAGATGGCTCTTAC TTCGTCTACA GCAAGCTCAA TGTGCAGAAG 1201 AGCAACTGGG AGGCAGGAAATACTTTCACC TGCTCTGTGT TACATGAGGG 1251 CCTGCACAAC CACCATACTG AGAAGAGCCTCTCCCACTCT CCTGGTAAAT 1301 GA

The amino acid sequence of the Ab-A HC including signal peptide is:

(SEQ ID NO: 29) 1 METGLRWLLL VAVLKGVHCQ SLEESGGRLV TPGTPLTLTC TASGFSLSSY51 WMNWVRQAPG EGLEWIGTID SGGRTDYASW AKGRFTISRT STTMDLKMTS 101 LTTGDTARYFCARNWNLWGQ GTLVTVSSAS TKGPSVYPLA PGSAAQTNSM 151 VTLGCLVKGY FPEPVTVTWNSGSLSSGVHT FPAVLQSDLY TLSSSVTVPS 201 STWPSETVTC NVAHPASSTK VDKKIVPRDCGCKPCICTVP EVSSVFIFPP 251 KPKDVLTITL TPKVTCVVVD ISKDDPEVQF SWFVDDVEVHTAQTQPREEQ 301 FNSTFRSVSE LPIMHQDWLN GKEFKCRVNS AAFPAPIEKT ISKTKGRPKA351 PQVYTIPPPK EQMAKDKVSL TCMITDFFPE DITVEWQWNG QPAENYKNTQ 401PIMNTNGSYF VYSKLNVQKS NWEAGNTFTC SVLHEGLHNH HTEKSLSHSP 451 GK

The nucleic acid sequence of Ab-A HC including signal peptide encodingsequence:

(SEQ ID NO: 30) 1 ATGGAGACTG GGCTGCGCTG GCTTCTCCTG GTCGCTGTGC TCAAAGGTGT51 CCACTGTCAG TCGCTGGAGG AGTCCGGGGG TCGCCTGGTC ACGCCTGGGA 101 CACCCCTGACACTCACCTGC ACAGCCTCTG GATTCTCCCT CAGTAGTTAT 151 TGGATGAACT GGGTCCGCCAGGCTCCAGGG GAGGGGCTGG AATGGATCGG 201 AACCATTGAT TCTGGTGGTA GGACGGACTACGCGAGCTGG GCAAAAGGCC 251 GATTCACCAT CTCCAGAACC TCGACTACGA TGGATCTGAAAATGACCAGT 301 CTGACGACCG GGGACACGGC CCGTTATTTC TGTGCCAGAA ATTGGAACTT351 GTGGGGCCAA GGCACCCTCG TCACCGTCTC GAGCGCTTCT ACAAAGGGCC 401CATCTGTCTA TCCACTGGCC CCTGGATCTG CTGCCCAAAC TAACTCCATG 451 GTGACCCTGGGATGCCTGGT CAAGGGCTAT TTCCCTGAGC CAGTGACAGT 501 GACCTGGAAC TCTGGATCCCTGTCCAGCGG TGTGCACACC TTCCCAGCTG 551 TCCTGCAGTC TGACCTCTAC ACTCTGAGCAGCTCAGTGAC TGTCCCCTCC 601 AGCACCTGGC CCAGCGAGAC CGTCACCTGC AACGTTGCCCACCCGGCCAG 651 CAGCACCAAG GTGGACAAGA AAATTGTGCC CAGGGATTGT GGTTGTAAGC701 CTTGCATATG TACAGTCCCA GAAGTATCAT CTGTCTTCAT CTTCCCCCCA 751AAGCCCAAGG ATGTGCTCAC CATTACTCTG ACTCCTAAGG TCACGTGTGT 801 TGTGGTAGACATCAGCAAGG ATGATCCCGA GGTCCAGTTC AGCTGGTTTG 851 TAGATGATGT GGAGGTGCACACAGCTCAGA CGCAACCCCG GGAGGAGCAG 901 TTCAACAGCA CTTTCCGCTC AGTCAGTGAACTTCCCATCA TGCACCAGGA 951 CTGGCTCAAT GGCAAGGAGT TCAAATGCAG GGTCAACAGTGCAGCTTTCC 1001 CTGCCCCCAT CGAGAAAACC ATCTCCAAAA CCAAAGGCAG ACCGAAGGCT1051 CCACAGGTGT ACACCATTCC ACCTCCCAAG GAGCAGATGG CCAAGGATAA 1101AGTCAGTCTG ACCTGCATGA TAACAGACTT CTTCCCTGAA GACATTACTG 1151 TGGAGTGGCAGTGGAATGGG CAGCCAGCGG AGAACTACAA GAACACTCAG 1201 CCCATCATGG ACACAGATGGCTCTTACTTC GTCTACAGCA AGCTCAATGT 1251 GCAGAAGAGC AACTGGGAGG CAGGAAATACTTTCACCTGC TCTGTGTTAC 1301 ATGAGGGCCT GCACAACCAC CATACTGAGA AGAGCCTCTCCCACTCTCCT 1351 GGTAAATGA

The CDR (complementarity determining region) sequences in the variableregion of the heavy chain of Ab-A are as follows:

(SEQ ID NO: 51) CDR-H1: SYWMN (SEQ ID NO: 52) CDR-H2: TIDSGGRTDYASWAKG(SEQ ID NO: 53) CDR-H3: NWNL

-   -   The light chain variable region CDR sequences of Ab-A are:

(SEQ ID NO: 54) CDR-L1: QSSQSVYDNNWLA (SEQ ID NO: 55) CDR-L2: DASDLAS(SEQ ID NO: 56) CDR-L3: QGAYNDVIYA

Ab-A was humanized, and is referred to as Antibody 1 (also referred toherein as Ab-1), having the following sequences:

The nucleic acid sequence of the Ab-1 LC variable region includingsignal peptide encoding sequence is

(SEQ ID NO: 74) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCTCAAGTTCTGACCCAGAGTCCAAGCAGTCTCTCCGCCAGCGTAGGCGATCGTGTGACTATTACCTGTCAATCTAGTCAGAGCGTGTATGATAACAATTGGCTGGCGTGGTACCAGCAAAAACCGGGCAAAGCCCCGAAGCTGCTCATCTATGACGCGTCCGATCTGGCTAGCGGTGTGCCAAGCCGTTTCAGTGGCAGTGGCAGCGGTACTGACTTTACCCTCACAATTTCGTCTCTCCAGCCGGAAGATTTCGCCACTTACTATTGTCAAGGTGCTTACAACGATGTGATTTATGCCTTCGGTCAGGGCACTAAAGTAGAAATCAAACGT

The amino acid sequence of Ab-1 LC variable region including signalpeptide is:

(SEQ ID NO: 75)

The nucleic acid sequence of Ab-1 HC variable region including signalpeptide encoding sequence is:

(SEQ ID NO: 76) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCACTGTGAGGTGCAGCTGTTGGAGTCTGGAGGCGGGCTTGTCCAGCCTGGAGGGAGCCTGCGTCTCTCTTGTGCAGCAAGCGGCTTCAGCTTATCCTCTTACTGGATGAATTGGGTGCGGCAGGCACCTGGGAAGGGCCTGGAGTGGGTGGGCACCATTGATTCCGGAGGCCGTACAGACTACGCGTCTTGGGCAAAGGGCCGTTTCACCATTTCCCGCGACAACTCCAAAAATACCATGTACCTCCAGATGAACTCTCTCCGCGCAGAGGACACAGCACGTTATTACTGTGCACGCAACTGGAATCTGTGGGGTCAAGGTACTCTTGTAACAGTCTCGAGCAmino acid sequence of Ab-1 HC variable region including signal peptide

(SEQ ID NO: 77)

The CDR (complementarity determining region) sequences in the variableregion of the heavy chain of Ab-1 are as follows:

(SEQ ID NO: 51) CDR-H1: SYWMN (SEQ ID NO: 52) CDR-H2: TIDSGGRTDYASWAKG(SEQ ID NO: 53) CDR-H3: NWNL

-   -   The light chain variable region CDR sequences of Ab-1 are:

(SEQ ID NO: 54) CDR-L1: QSSQSVYDNNWLA (SEQ ID NO: 55) CDR-L2: DASDLAS(SEQ ID NO: 56) CDR-L3: QGAYNDVIYA

Ab-B

Antibody B (also referred to herein as Ab-B and Mab-B) is a mouseantibody which exhibits high affinity binding to sclerostin. The BIAcorebinding pattern of Ab-B is shown in FIG. 16.

Ab-B Light Chain

The amino acid sequence of the mature form (signal peptide removed) ofthe Ab-B LC is:

(SEQ ID NO: 31)

The nucleic acid sequence encoding the mature form (signal peptideremoved) of Ab-B LC is:

(SEQ ID NO: 32) 1 CAAATTGTTC TCACCCAGTC TCCAACAATC GTGTCTGCAT CTCCAGGGGA51 GAAGGTCACC CTAATCTGCA GTGCCAGTTC AAGTGTAAGT TTCGTGGACT 101 GGTTCCAGCAGAAGGCAGGC ACTTCTCCCA AACGCTGGAT TTACAGAACA 151 TCCAACCTGG GTTTTGGAGTCCCTGCTCGC TTCAGTGGCG GTGGATCTGG 201 GACCTCTCAC TCTCTCACAA TCAGCCGAATGGAGGCTGAA GATGCTGCCA 251 CTTATTACTG CCAGCAAAGG AGTACTTACC CACCCACGTTCGGTGCTGGG 301 ACCAAGCTGG AACTGAAACG GGCTGATGCT GCACCAACTG TATCCATCTT351 CCCACCATCC AGTGAGCAGT TAACATCTGG AGGTGCCTCA GTCGTGTGCT 401TCTTGAACAA CTTCTACCCC AAAGACATCA ATGTCAAGTG GAAGATTGAT 451 GGCAGTGAACGACAAAATGG CGTCCTGAAC AGTTGGACTG ATCAGGACAG 501 CAAAGACAGC ACCTACAGCATGAGCAGCAC CCTCACGTTG ACCAAGGACG 551 AGTATGAACG ACATAACAGC TATACCTGTGAGGCCACTCA CAAGAGATCA 601 ACTTCACCCA TTGTCAAGAG CTTCAACAGG AATGAGTGTT AG

The amino acid sequence of Ab-B LC including signal peptide is:

(SEQ ID NO: 33) 1 MHFQVQIFSF LLISASVIVS RGQIVLTQSP TIVSASPGEK VTLICSASSS51 VSFVDWFQQK PGTSPKRWIY RTSNLGFGVP ARFSGGGSGT SHSLTISRME 101 AEDAATYYCQQRSTYPPTFG AGTKLELKRA DAAPTVSIFP PSSEQLTSGG 151 ASVVCFLNNF YPKDINVKWKIDGSERQNGV LNSWTDQDSK DSTYSMSSTL 201 TLTKDEYERH NSYTCEATHK TSTSPIVKSFNRNEC

The nucleic acid sequence of Ab-B LC including signal peptide encodingsequence is:

(SEQ ID NO: 34) 1 ATGCATTTTC AAGTGCAGAT TTTCAGCTTC CTGCTAATCA GTGCCTCAGT51 CATAGTGTCC AGAGGGCAAA TTGTTCTCAC CCAGTCTCCA ACAATCGTGT 101 CTGCATCTCCAGGGGAGAAG GTCACCCTAA TCTGCAGTGC CAGTTCAAGT 151 GTAAGTTTCG TGGACTGGTTCCAGCAGAAG CCAGGCACTT CTCCCAAACG 201 CTGGATTTAC AGAACATCCA ACCTGGGTTTTGGAGTCCCT GCTCGCTTCA 251 GTGGCGGTGG ATCTGGGACC TCTCACTCTC TCACAATCAGCCGAATGGAG 301 GCTGAAGATG CTGCCACTTA TTACTGCCAG CAAAGGAGTA CTTACCCACC351 CACGTTCGGT GCTGGGACCA AGCTGGAACT GAAACGGGCT GATGCTGCAC 401CAACTGTATC CATCTTCCCA CCATCCAGTG AGCAGTTAAC ATCTGGAGGT 451 GCCTCAGTCGTGTGCTTCTT GAACAACTTC TACCCCAAAG ACATCAATGT 501 CAAGTGGAAG ATTGATGGCAGTGAACGACA AAATGGCGTC CTGAACAGTT 551 GGACTGATCA GGACAGCAAA GACAGCACCTACAGCATGAG CAGCACCCTC 601 ACGTTGACCA AGGACGAGTA TGAACGACAT AACAGCTATACCTGTGAGGC 651 CACTCACAAG ACATCAACTT CACCCATTGT CAAGAGCTTC AACAGGAATG701 AGTGTTAG

Ab-B Heavy Chain

The amino acid sequence of the mature form (signal peptide removed) ofAb-B HC:

(SEQ ID NO: 35)

The nucleic acid sequence encoding the mature form (signal peptideremoved) of Ab-B HC:

(SEQ ID NO: 36) 1 CAGGTTACTC TGAAAGAGTC TGGCCCTGGG ATATTGCAGC CCTCCCAGAC51 CCTCAGTCTG ACTTGTTCTT TCTCTGGGTT TTCACTGAGC ACTTCTGGTA 101 TGGGTGTAGGCTGGATTCGT CACCCATCAG GGAAGAATCT GGAGTGGCTG 151 GCACACATTT GGTGGGATGATGTCAAGCGC TATAACCCAG TCCTGAAGAG 201 CCGACTGACT ATCTCCAAGG ATACCTCCAACAGCCAGGTA TTCCTCAAGA 251 TCGCCAATGT GGACACTGCA GATACTGCCA CATACTACTGTGCTCGAATA 301 GAGGACTTTG ATTACGACGA GGAGTATTAT GCTATGGACT ACTGGGGTCA351 AGGAACCTCA GTCATCGTCT CCTCAGCCAA AACGACACCC CCATCTGTCT 401ATCCACTGGC CCCTGGATCT GCTGCCCAAA CTAACTCCAT GGTGACCCTG 451 GGATGCCTGGTCAAGGGCTA TTTCCCTGAG CCAGTGACAG TGACCTGGAA 501 CTCTGGATCC CTGTCCAGCGGTGTGCACAC CTTCCCAGCT GTCCTGCAGT 551 CTGACCTCTA CACTCTGAGC AGCTCAGTGACTGTCCCCTC CAGCACCTGG 601 CCCAGCGAGA CCGTCACCTG CAACGTTGCC CACCCGGCCAGCAGCACCAA 651 GGTGGACAAG AAAATTGTGC CCAGGGATTG TGGTTGTAAG CCTTGCATAT701 GTACAGTCCC AGAAGTATCA TCTGTCTTCA TCTTCCCCCC AAAGCCCAAG 751GATGTGCTCA CCATTACTCT GACTCCTAAG GTCACGTGTG TTGTGGTAGA 801 CATCAGCAAGGATGATCCCG AGGTCCAGTT CAGCTGGTTT GTAGATGATG 851 TGGAGGTGCA CACAGCTCAGACGCAACCCC GGGAGGAGCA GTTCAACAGC 901 ACTTTCCGCT CAGTCAGTGA ACTTCCCATCATGCACCAGG ACTGGCTCAA 951 TGGCAAGGAG TTCAAATGCA GGGTCAACAG TGCAGCTTTCCCTGCCCCCA 1001 TCGAGAAAAC CATCTCCAAA ACCAAAGGCA GACCGAAGGC TCCACAGGTG1051 TACACCATTC CACCTCCCAA GGAGCAGATG GCCAAGGATA AAGTCAGTCT 1101GACCTGCATG ATAACAGACT TCTTCCCTGA AGACATTACT GTGGAGTGGC 1151 AGTGGAATGGGCAGCCAGCG GAGAACTACA AGAACACTCA GCCCATCATG 1201 GACACAGATG GCTCTTACTTCGTCTACAGC AAGCTCAATG TGCAGAAGAG 1251 CAACTGGGAG GCAGGAAATA CTTTCACCTGCTCTGTGTTA CATGAGGGCC 1301 TGCACAACCA CCATACTGAG AAGAGCCTCT CCCACTCTCCTGGTAAATGA

The amino acid sequence of Ab-B HC including signal peptide:

(SEQ ID NO: 37) 1 MGRLTSSFLL LIVPAYVLSQ VTLKESGPGI LQPSQTLSLT CSFSGFSLST51 SGMGVGWIRH PSGKNLEWLA HIWWDDVKRY NPVLKSRLTI SKDTSNSQVF 101 LKIANVDTADTATYYCARIE DFDYDEEYYA MDYWGQGTSV IVSSAKTTPP 151 SVYPLAPGSA AQTNSMVTLGCLVKGYFPEP VTVTWNSGSL SSGVHTFPAV 201 LQSDLYTLSS SVTVPSSTWP SETVTCNVAHPASSTKVDKK IVPRDCGCKP 251 CICTVPEVSS VFIFPPKPKD VLTITLTPKV TCVVVDISKDDPEVQFSWFV 301 DDVEVHTAQT QPREEQFNST FRSVSELPIM HQDWLNGKEF KCRVNSAAFP351 APIEKTISKT KGRPKAPQVY TIPPPKEQMA KDKVSLTCMI TDFFPEDITV 401EWQWNGQPAE NYKNTQPIMD TDGSYFVYSK LNVQKSNWEA GNTFTCSVLH 451 EGLHNHHTEKSLSHSPGK

The nucleic acid sequence of Ab-B HC including signal peptide encodingsequence:

(SEQ ID NO: 38) 1 ATGGGCAGGC TTACTTCTTC ATTCCTGCTA CTGATTGTCC CTGCATATGT51 CCTGTCCCAG GTTACTCTGA AAGAGTCTGG CCCTGGGATA TTGCAGCCCT 101 CCCAGACCCTCAGTCTGACT TGTTCTTTCT CTGGGTTTTC ACTGAGCACT 151 TCTGGTATGG GTGTAGGCTGGATTCGTCAC CCATCAGGGA AGAATCTGGA 201 GTGGCTGGCA CACATTTGGT GGGATGATGTCAAGCGCTAT AACCCAGTCC 251 TGAAGAGCCG ACTGACTATC TCCAAGGATA CCTCCAACAGCCAGGTATTC 301 CTCAAGATCG CCAATGTGGA CACTGCAGAT ACTGCCACAT ACTACTGTGC351 TCGAATAGAG GACTTTGATT ACGACGAGGA GTATTATGCT ATGGACTACT 401GGGGTCAAGG AACCTCAGTC ATCGTCTCCT CAGCCAAAAC GACACCCCCA 451 TCTGTCTATCCACTGGCCCC TGGATCTGCT GCCCAAACTA ACTCCATGGT 501 GACCCTGGGA TGCCTGGTCAAGGGCTATTT CCCTGAGCCA GTGACAGTGA 551 CCTGGAACTC TGGATCCCTG TCCAGCGGTGTGCACACCTT CCCAGCTGTC 601 CTGCAGTCTG ACCTCTACAC TCTGAGCAGC TCAGTGACTGTCCCCTCCAG 651 CACCTGGCCC AGCGAGACCG TCACCTGCAA CGTTGCCCAC CCGGCCAGCA701 GCACCAAGGT GGACAAGAAA ATTGTGCCCA GGGATTGTGG TTGTAAGCCT 751TGCATATGTA CAGTCCCAGA AGTATCATCT GTCTTCATCT TCCCCCCAAA 801 GCCCAAGGATGTGCTCACCA TTACTCTGAC TCCTAAGGTC ACGTGTGTTG 851 TGGTAGACAT CAGCAAGGATGATCCCGAGG TCCAGTTCAG CTGGTTTGTA 901 GATGATGTGG AGGTGCACAC AGCTCAGACGCAACCCCGGG AGGAGCAGTT 951 CAACAGCACT TTCCGCTCAG TCAGTGAACT TCCCATCATGCACCAGGACT 1001 GGCTCAATGG CAAGGAGTTC AAATGCAGGG TCAACAGTGC AGCTTTCCCT1051 GCCCCCATCG AGAAAACCAT CTCCAAAACC AAAGGCAGAC CGAAGGCTCC 1101ACAGGTGTAC ACCATTCCAC CTCCCAAGGA GCAGATGGCC AAGGATAAAG 1151 TCAGTCTGACCTGCATGATA ACAGACTTCT TCCCTGAAGA CATTACTGTG 1201 GAGTGGCAGT GGAATGGGCAGCCAGCGGAG AACTACAAGA ACACTCAGCC 1251 CATCATGGAC ACAGATGGCT CTTACTTCGTCTACAGCAAG CTCAATGTGC 1301 AGAAGAGCAA CTGGGAGGCA GGAAATACTT TCACCTGCTCTGTGTTACAT 1351 GAGGGCCTGC ACAACCACCA TACTGAGAAG AGCCTCTCCC ACTCTCCTGG1401 TAAATGA

The CDR (complementarity determining region) sequences in the variableregion of the heavy chain of Ab-B are as follows:

(SEQ ID NO: 57) CDR-H1: TSGMGVG (SEQ ID NO: 58) CDR-H2: HIWWDDVKRYNPVLKS(SEQ ID NO: 59) CDR-H3: EDFDYDEEYYAMDY

-   -   The light chain variable region CDR sequences of Ab-B are:

(SEQ ID NO: 60) CDR-L1: SASSSVSFVD (SEQ ID NO: 61) CDR-L2: RTSNLGF (SEQID NO: 62) CDR-L3: QQRSTYPPT

Antibodies disclosed herein bind to regions of human sclerostin whichare important for the in vivo activity of the protein. Binding of anantibody to sclerostin can be correlated with increases in, for example,the bone mineral density achieved by use of the antibody in vivo such asdescribed in Examples 5 and 9 (mice) and Example 12 (monkey). Increasesin at least one of bone formation, bone mineral content, bone mass, bonequality and bone strength can also be achieved by use of the antibody invivo such as described in Examples 5 and 9 (mice) and Example 12(monkey). Since the binding of an antibody to sclerostin is primarilydetermined by its CDR sequences, an antibody for practicing theinvention may be generated with all or some of the disclosed CDRsequences in an appropriate framework, wherein the antibody retains theability to bind specifically to sclerostin, and can be expected toachieve increases in, for example, bone mineral density. Such antibodiesare useful in the treatment of human or animal conditions that arecaused by, associated with, or result in at least one of low boneformation, low bone mineral density, low bone mineral content, low bonemass, low bone quality and low bone strength. Methods of constructingand expressing antibodies and fragments thereof comprising CDR's of thepresent invention are known to those of skill in the art.

The present invention therefore relates in one embodiment to an isolatedantibody, including Ab-A, or an antigen binding fragment thereof, whichspecifically binds to sclerostin and wherein the variable domain of theheavy chain comprises at least one CDR having the sequences given in SEQID NO:51 for CDR-H1, SEQ ID NO:52 for CDR-H2 and SEQ ID NO:53 forCDR-H3. The antibody or antigen binding fragment thereof may comprise aheavy chain variable domain in which the CDRs consist of at least one ofthe peptides of SEQ ID NO:51 for CDR-H1, SEQ ID NO:52 for CDR-H2 and SEQID NO:53 for CDR-H3.

When in antibodies of the invention a light chain is present the lightchain may be any suitable complementary chain and may in particular beselected from a light chain wherein the variable domain comprises atleast one CDR having the sequences given in SEQ ID NO:54 for CDR-L1, SEQID NO:55 for CDR-L2 and SEQ ID NO:56 for CDR-L3. The antibody or antigenbinding fragment thereof may comprise a light chain variable domain inwhich the CDRs consist of at least one of the peptides of SEQ ID NO:54for CDR-L1, SEQ ID NO:55 for CDR-L2 and SEQ ID NO:56 for CDR-L3.

-   -   The present invention further relates to an isolated antibody,        including Ab-B, or an antigen binding fragment hereof, which        specifically binds to sclerostin and wherein the variable domain        of the heavy chain comprises at least one CDR having the        sequences given in SEQ ID NO:57 for CDR-H1, SEQ ID NO:58 for        CDR-H2 and SEQ ID NO:59 for CDR-H3. The antibody or antigen        binding fragment thereof may comprise a heavy chain variable        domain in which the CDRs consist of at least one of the peptides        of SEQ ID NO:57 for CDR-H1, SEQ ID NO:58 for CDR-H2 and SEQ ID        NO:59 for CDR-H3.

When in antibodies of the invention a light chain is present the lightchain may be any suitable complementary chain and may in particular beselected from a light chain wherein the variable domain comprises atleast one CDR having the sequences given in SEQ ID NO:60 for CDR-L1, SEQID NO:61 for CDR-L2 and SEQ ID NO:62 for CDR-L3. The antibody or antigenbinding fragment thereof may comprise a light chain variable domain inwhich the CDRs consist of at least one of the peptides of SEQ ID NO:60for CDR-L1, SEQ ID NO:61 for CDR-L2 and SEQ ID NO:62 for CDR-L3.

The present invention still further relates to an isolated antibody,including Ab-C, or an antigen binding fragment hereof, whichspecifically binds to sclerostin and wherein the variable domain of theheavy chain comprises at least one CDR having the sequences given in SEQID NO:45 for CDR-H1, SEQ ID NO:46 for CDR-H2 and SEQ ID NO:47 forCDR-H3. The antibody or antigen binding fragment thereof may comprise aheavy chain variable domain in which the CDRs consist of at least one ofthe peptides of SEQ ID NO:45 for CDR-H1, SEQ ID NO:46 for CDR-H2 and SEQID NO:47 for CDR-H3.

When in antibodies of the invention a light chain is present the lightchain may be any suitable complementary chain and may in particular beselected from a light chain wherein the variable domain comprises atleast one CDR having the sequences given in SEQ ID NO:48 for CDR-L1, SEQID NO:49 for CDR-L2 and SEQ ID NO:50 for CDR-L3. The antibody or antigenbinding fragment thereof may comprise a light chain variable domain inwhich the CDRs consist of at least one of the peptides of SEQ ID NO:48for CDR-L1, SEQ ID NO:49 for CDR-L2 and SEQ ID NO:50 for CDR-L3.

The present invention also relates to an isolated antibody, includingAb-D, or an antigen binding fragment hereof, which specifically binds tosclerostin and wherein the variable domain of the heavy chain comprisesat least one CDR having the sequences given in SEQ ID NO:39 for CDR-H1,SEQ ID NO:40 for CDR-H2 and SEQ ID NO:41 for CDR-H3. The antibody orantigen binding fragment thereof may comprise a heavy chain variabledomain in which the CDRs consist of at least one of the peptides of SEQID NO:39 for CDR-H1, SEQ ID NO:40 for CDR-H2 and SEQ ID NO:41 forCDR-H3.

When in antibodies of the invention a light chain is present the lightchain may be any suitable complementary chain and may in particular beselected from a light chain wherein the variable domain comprises atleast one CDR having the sequences given in SEQ ID NO:42 for CDR-L1, SEQID NO:43 for CDR-L2 and SEQ ID NO:44 for CDR-L3. The antibody or antigenbinding fragment thereof may comprise a light chain variable domain inwhich the CDRs consist of at least one of the peptides of SEQ ID NO:42for CDR-L1, SEQ ID NO:43 for CDR-L2 and SEQ ID NO:44 for CDR-L3.

Additional anti-sclerostin antibodies are described below. For some ofthe amino acid sequences the complementarity-determining regions (CDRs)are boxed-shaded and the constant regions are in bold-italics.

Ab-2

The sequences of the Antibody 2 (also referred to as Ab-2) LC and HC areas follows:

Ab-2 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-2 LC:

(SEQ ID NO: 117)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-2 LC:

(SEQ ID NO: 118) 1 CAAATTGTTC TCTCCCAGTC TCCAGCAATC CTGTCTACATCTCCAGGGGA 51 GAAGGTCACA ATGACTTGCA GGGCCAGCTC AAGTGTATAT TACATGCACT 101GGTACCAGCA GAAGCCAGGA TCCTCCCCCA AACCCTGGAT TTATGCCACA 151 TCCAACCTGGCTTCTGGAGT CCCTGTTCGC TTCAGTGGCA GTGGGTCTGG 201 GACCTCTTAC TCTCTCACAATCACCAGAGT GGAGGCTGAA GATGCTGCCA 251 CTTATTACTG CCAGCAGTGG AGTAGTGACCCACTCACGTT CGGTGCTGGG 301 ACCAAGCTGG AGCTGAAACG GGCTGATGCT GCACCAACTGTATCCATCTT 351 CCCACCATCC AGTGAGCAGT TAACATCTGG AGGTGCCTCA GTCGTGTGCT401 TCTTGAACAA CTTCTACCCC AAAGACATCA ATGTCAAGTG GAAGATTGAT 451GGCAGTGAAC GACAAAATGG CGTCCTGAAC AGTTGGACTG ATCAGGACAG 501 CAAAGACAGCACCTACAGCA TGAGCAGCAC CCTCACGTTG ACCAAGGACG 551 AGTATGAACG ACATAACAGCTATACCTGTG AGGCCACTCA CAAGACATCA 601 ACTTCACCCA TTGTCAAGAG CTTCAACAGGAATGAGTGTT AGAmino acid sequence of the Ab-2 LC including signal peptide:

(SEQ ID NO: 119) 1 MDFQVQIFSF LLISASVIMS RGQIVLSQSP AILSTSPGEKVTMTCRASSS 51 VYYMHWYQQK PGSSPKPWIY ATSNLASGVP VRFSGSGSGT SYSLTITRVE 101AEDAATYYCQ QWSSDPLTFG AGTKLELKRA DAAPTVSIFP PSSEQLTSGG 151 ASVVCFLNNFYPKDINVKWK IDGSERQNGV LNSWTDQDSK DSTYSMSSTL 201 TLTKDEYERH NSYTCEATHKTSTSPIVKSF NRNECNucleic acid sequence of the Ab-2 LC including signal peptide encodingsequence:

(SEQ ID NO: 120) 1 ATGGATTTTC AAGTGCAGAT TTTCAGCTTC CTGCTAATCAGTGCTTCAGT 51 CATTATGTCC AGGGGACAAA TTGTTCTCTC CCAGTCTCCA GCAATCCTGT 101CTACATCTCC AGGGGAGAAG GTCACAATGA CTTGCAGGGC CAGCTCAAGT 151 GTATATTACATGCACTGGTA CCAGCAGAAG CCAGGATCCT CCCCCAAACC 201 CTGGATTTAT GCCACATCCAACCTGGCTTC TGGAGTCCCT GTTCGCTTCA 251 GTGGCAGTGG GTCTGGGACC TCTTACTCTCTCACAATCAC CAGAGTGGAG 301 GCTGAAGATG CTGCCACTTA TTACTGCCAG CAGTGGAGTAGTGACCCACT 351 CACGTTCGGT GCTGGGACCA AGCTGGAGCT GAAACGGGCT GATGCTGCAC401 CAACTGTATC CATCTTCCCA CCATCCAGTG AGCAGTTAAC ATCTGGAGGT 451GCCTCAGTCG TGTGCTTCTT GAACAACTTC TACCCCAAAG ACATCAATGT 501 CAAGTGGAAGATTGATGGCA GTGAACGACA AAATGGCGTC CTGAACAGTT 551 GGACTGATCA GGACAGCAAAGACAGCACCT ACAGCATGAG CAGCACCCTC 601 ACGTTGACCA AGGACGAGTA TGAACGACATAACAGCTATA CCTGTGAGGC 651 CACTCACAAG ACATCAACTT CACCCATTGT CAAGAGCTTCAACAGGAATG 701 AGTGTTAG

Ab-2 Heavy Chain

Amino acid sequence of the mature form (signal peptide removed) of theAb-2 HC:

(SEQ ID NO: 121)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-2 HC:

(SEQ ID NO: 122) 1 GAGGTTCAGG TGCAGCAGTC TGGGCCAGAA CTTGTGAAGCCAGGGGCCTC 51 AGTCAAGTTG TCCTGCACAG CTTCTGGCTT CAACATTAAA GACTACTTTA 101TACACTGGGT GAAGCAGAGG CCTGAACAGG GCCTGGAGTG GATTGGAAGG 151 CTTGATCCTGAGGATGGTGA AAGTGATTAT GCCCCGAAGT TCCAGGACAA 201 GGCCATTATG ACAGCAGACACATCATCCAA CACAGCCTAT CTTCAGCTCA 251 GAAGCCTGAC ATCTGAGGAC ACTGCCATCTATTATTGTGA GAGAGAGGAC 301 TACGATGGTA CCTACACCTT TTTTCCTTAC TGGGGCCAAGGGACTCTGGT 351 CACTGTCTCT GCAGCCAAAA CGACACCCCC ATCTGTCTAT CCACTGGCCC401 CTGGATCTGC TGCCCAAACT AACTCCATGG TGACCCTGGG ATGCCTGGTC 451AAGGGCTATT TCCCTGAGCC AGTGACAGTG ACCTGGAACT CTGGATCCCT 501 GTCCAGCGGTGTGCACACCT TCCCAGCTGT CCTGCAGTCT GACCTCTACA 551 CTCTGAGCAG CTCAGTGACTGTCCCCTCCA GCACCTGGCC CAGCGAGACC 601 GTCACCTGCA ACGTTGCCCA CCCGGCCAGCAGCACCAAGG TGGACAAGAA 651 AATTGTGCCC AGGGATTGTG GTTGTAAGCC TTGCATATGTACAGTCCCAG 701 AAGTATCATC TGTCTTCATC TTCCCCCCAA AGCCCAAGGA TGTGCTCACC751 ATTACTCTGA CTCCTAAGGT CACGTGTGTT GTGGTAGACA TCAGCAAGGA 801TGATCCCGAG GTCCAGTTCA GCTGGTTTGT AGATGATGTG GAGGTGCACA 851 CAGCTCAGACGCAACCCCGG GAGGAGCAGT TCAACAGCAC TTTCCGCTCA 901 GTCAGTGAAC TTCCCATCATGCACCAGGAC TGGCTCAATG GCAAGGAGTT 951 CAAATGCAGG GTCAACAGTG CAGCTTTCCCTGCCCCCATC GAGAAAACCA 1001 TCTCCAAAAC CAAAGGCAGA CCGAAGGCTC CACAGGTGTACACCATTCCA 1051 CCTCCCAAGG AGCAGATGGC CAAGGATAAA GTCAGTCTGA CCTGCATGAT1101 AACAGACTTC TTCCCTGAAG ACATTACTGT GGAGTGGCAG TGGAATGGGC 1151AGCCAGCGGA GAACTACAAG AACACTCAGC CCATCATGGA CACAGATGGC 1201 TCTTACTTCATCTACAGCAA GCTCAATGTG CAGAAGAGCA ACTGGGAGGC 1251 AGGAAATACT TTCACCTGCTCTGTGTTACA TGAGGGCCTG CACAACCACC 1301 ATACTGAGAA GAGCCTCTCC CACTCTCCTGGTAAATGAAmino acid sequence of the Ab-2 HC including signal peptide:

(SEQ ID NO: 123) 1 MKCSWVIFFL MAVVTGVNSE VQVQQSGPEL VKPGASVKLSCTASGFNIKD 51 YFIHWVKQRP EQGLEWIGRL DPEDGESDYA PKFQDKAIMT ADTSSNTAYL 101QLRSLTSEDT AIYYCEREDY DGTYTFFPYW GQGTLVTVSA AKTTPPSVYP 151 LAPGSAAQTNSMVTLGCLVK GYFPEPVTVT WNSGSLSSGV HTFPAVLQSD 201 LYTLSSSVTV PSSTWPSETVTCNVAHPASS TKVDKKIVPR DCGCKPCICT 251 VPEVSSVFIF PPKPKDVLTI TLTPKVTCVVVDISKDDPEV QFSWFVDDVE 301 VHTAQTQPRE EQFNSTFRSV SELPIMHQDW LNGKEFKCRVNSAAFPAPIE 351 KTISKTKGRP KAPQVYTIPP PKEQMAKDKV SLTCMITDFF PEDITVEWQW401 NGQPAENYKN TQPIMDTDGS YFIYSKLNVQ KSNWEAGNTF TCSVLHEGLH 451NHHTEKSLSH SPGKNucleic acid sequence of the Ab-2 HC including signal peptide encodingsentience:

(SEQ ID NO: 124) 1 ATGAAATGCA GCTGGGTCAT CTTCTTCCTG ATGGCAGTGGTTACAGGGGT 51 CAATTCAGAG GTTCAGGTGC AGCAGTCTGG GCCAGAACTT GTGAAGCCAG 101GGGCCTCAGT CAAGTTGTCC TGCACAGCTT CTGGCTTCAA CATTAAAGAC 151 TACTTTATACACTGGGTGAA GCAGAGGCCT GAACAGGGCC TGGAGTGGAT 201 TGGAAGGCTT GATCCTGAGGATGGTGAAAG TGATTATGCC CCGAAGTTCC 251 AGGACAAGGC CATTATGACA GCAGACACATCATCCAACAC AGCCTATCTT 301 CAGCTCAGAA GCCTGACATC TGAGGACACT GCCATCTATTATTGTGAGAG 351 AGAGGACTAC GATGGTACCT ACACCTTTTT TCCTTACTGG GGCCAAGGGA401 CTCTGGTCAC TGTCTCTGCA GCCAAAACGA CACCCCCATC TGTCTATCCA 451CTGGCCCCTG GATCTGCTGC CCAAACTAAC TCCATGGTGA CCCTGGGATG 501 CCTGGTCAAGGGCTATTTCC CTGAGCCAGT GACAGTGACC TGGAACTCTG 551 GATCCCTGTC CAGCGGTGTGCACACCTTCC CAGCTGTCCT GCAGTCTGAC 601 CTCTACACTC TGAGCAGCTC AGTGACTGTCCCCTCCAGCA CCTGGCCCAG 651 CGAGACCGTC ACCTGCAACG TTGCCCACCC GGCCAGCAGCACCAAGGTGG 701 ACAAGAAAAT TGTGCCCAGG GATTGTGGTT GTAAGCCTTG CATATGTACA751 GTCCCAGAAG TATCATCTGT CTTCATCTTC CCCCCAAAGC CCAAGGATGT 801GCTCACCATT ACTCTGACTC CTAAGGTCAC GTGTGTTGTG GTAGACATCA 851 GCAAGGATGATCCCGAGGTC CAGTTCAGCT GGTTTGTAGA TGATGTGGAG 901 GTGCACACAG CTCAGACGCAACCCCGGGAG GAGCAGTTCA ACAGCACTTT 951 CCGCTCAGTC AGTGAACTTC CCATCATGCACCAGGACTGG CTCAATGGCA 1001 AGGAGTTCAA ATGCAGGGTC AACAGTGCAG CTTTCCCTGCCCCCATCGAG 1051 AAAACCATCT CCAAAACCAA AGGCAGACCG AAGGCTCCAC AGGTGTACAC1101 CATTCCACCT CCCAAGGAGC AGATGGCCAA GGATAAAGTC AGTCTGACCT 1151GCATGATAAC AGACTTCTTC CCTGAAGACA TTACTGTGGA GTGGCAGTGG 1201 AATGGGCAGCCAGCGGAGAA CTACAAGAAC ACTCAGCCCA TCATGGACAC 1251 AGATGGCTCT TACTTCATCTACAGCAAGCT CAATGTGCAG AAGAGCAACT 1301 GGGAGGCAGG AAATACTTTC ACCTGCTCTGTGTTACATGA GGGCCTGCAC 1351 AACCACCATA CTGAGAAGAG CCTCTCCCAC TCTCCTGGTAAATGA

Ab-3

The sequences of the Antibody 3 (also referred to herein as Ab-3) LC andHC are as follows:

Ab-3 Light Chain

Amino acid sequence of the mature form (signal peptide removed) of theAb-3 LC:

(SEQ ID NO: 125)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-3 LC:

(SEQ ID NO: 126) 1 GAAATTGTGC TCACCCAGTC TCCAGCACTC ATGGCTGCATCTCCGGGGGA 51 GAAGGTCACC ATCACCTGCA GTGTCAGTTC AACTATAAGT TCCAACCACT 101TGCACTGGTT CCAGCAGAAG TCAGACACCT CCCCCAAACC CTGGATTTAT 151 GGCACATCCAACCTGGCTTC TGGAGTCCCT GTTCGCTTCA GTGGCAGTGG 201 ATCTGGGACC TCTTATTCTCTCACAATCAG CAGCATGGAG GCTGAGGATG 251 CTGCCACTTA TTACTGTCAA CAGTGGAGTAGTTACCCACT CACGTTCGGC 301 GCTGGGACCA AGCTGGAGCT GAGACGGGCT GATGCTGCACCAACTGTATC 351 CATCTTCCCA CCATCCAGTG AGCAGTTAAC ATCTGGAGGT GCCTCAGTCG401 TGTGCTTCTT GAACAACTTC TACCCCAAAG ACATCAATGT CAAGTGGAAG 451ATTGATGGCA GTGAACGACA AAATGGCGTC CTGAACAGTT GGACTGATCA 501 GGACAGCAAAGACAGCACCT ACAGCATGAG CAGCACCCTC ACGTTGACCA 551 AGGACGAGTA TGAACGACATAACAGCTATA CCTGTGAGGC CACTCACAAG 601 ACATCAACTT CACCCATTGT CAAGAGCTTCAACAGGAATG AGTGTTAGAmino acid sequence of the Ab-3 LC including signal peptide:

(SEQ ID NO: 127) 1 MDFHVQIFSF MLISVTVILS SGEIVLTQSP ALMAASPGEKVTITCSVSST 51 ISSNHLHWFQ QKSDTSPKPW IYGTSNLASG VPVRFSGSGS GTSYSLTISS 101MEAEDAATYY CQQWSSYPLT FGAGTKLELR RADAAPTVSI FPPSSEQLTS 151 GGASVVCFLNNFYPKDINVK WKIDGSERQN GVLNSWTDQD SKDSTYSMSS 201 TLTLTKDEYE RHNSYTCEATHKTSTSPIVK SFNRNECNucleic acid sequence of the Ab-3 LC including signal peptide encodingsequence:

(SEQ ID NO: 128) 1 ATGGATTTTC ATGTGCAGAT TTTCAGCTTC ATGCTAATCAGTGTCACAGT 51 CATTTTGTCC AGTGGAGAAA TTGTGCTCAC CCAGTCTCCA GCACTCATGG 101CTGCATCTCC GGGGGAGAAG GTCACCATCA CCTGCAGTGT CAGTTCAACT 151 ATAAGTTCCAACCACTTGCA CTGGTTCCAG CAGAAGTCAG ACACCTCCCC 201 CAAACCCTGG ATTTATGGCACATCCAACCT GGCTTCTGGA GTCCCTGTTC 251 GCTTCAGTGG CAGTGGATCT GGGACCTCTTATTCTCTCAC AATCAGCAGC 301 ATGGAGGCTG AGGATGCTGC CACTTATTAC TGTCAACAGTGGAGTAGTTA 351 CCCACTCACG TTCGGCGCTG GGACCAAGCT GGAGCTGAGA CGGGCTGATG401 CTGCACCAAC TGTATCCATC TTCCCACCAT CCAGTGAGCA GTTAACATCT 451GGAGGTGCCT CAGTCGTGTG CTTCTTGAAC AACTTCTACC CCAAAGACAT 501 CAATGTCAAGTGGAAGATTG ATGGCAGTGA ACGACAAAAT GGCGTCCTGA 551 ACAGTTGGAC TGATCAGGACAGCAAAGACA GCACCTACAG CATGAGCAGC 601 ACCCTCACGT TGACCAAGGA CGAGTATGAACGACATAACA GCTATACCTG 651 TGAGGCCACT CACAAGACAT CAACTTCACC CATTGTCAAGAGCTTCAACA 701 GGAATGAGTG TTAG

Ab-3 Heavy Chain

Amino acid sequence of the mature form (signal peptide removed) of theAb-3 HC:

(SEQ ID NO: 129)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-3 HC:

(SEQ ID NO: 130) 1 GAGGTTCAGC TGCAGCAGTC TGGGGCTGAA CTTGTGAGGCCAGGGGCCTT 51 AGTCAAGTTG TCCTGCACAG CTTCTGACTT CAACATTAAA GACTTCTATC 101TACACTGGAT GAGGCAGCGG CCTGAACAGG GCCTGGACTG GATTGGAAGG 151 ATTGATCCTGAGAATGGTGA TACTTTATAT GACCCGAAGT TCCAGGACAA 201 GGCCACTCTT ACAACAGACACATCCTCCAA CACAGCCTAC CTGCAGCTCA 251 GCGGCCTGAC ATCTGAGACC ACTGCCGTCTATTACTGTTC TAGAGAGGCG 301 GATTATTTCC ACGATGGTAC CTCCTACTGG TACTTCGATGTCTGGGGCGC 351 AGGGACCACA ATCACCGTCT CCTCAGCCAA AACGACACCC CCATCTGTCT401 ATCCACTGGC CCCTGGATCT GCTGCCCAAA CTAACTCCAT GGTGACCCTG 451GGATGCCTGG TCAAGGGCTA TTTCCCTGAG CCAGTGACAG TGACCTGGAA 501 CTCTGGATCCCTGTCCAGCG GTGTGCACAC CTTCCCAGCT GTCCTGCAGT 551 CTGACCTCTA CACTCTGAGCAGCTCAGTGA CTGTCCCCTC CAGCACCTGG 601 CCCAGCGAGA CCGTCACCTG CAACGTTGCCCACCCGGCCA GCAGCACCAA 651 GGTGGACAAG AAAATTGTGC CCAGGGATTG TGGTTGTAAGCCTTGCATAT 701 GTACAGTCCC AGAAGTATCA TCTGTCTTCA TCTTCCCCCC AAAGCCCAAG751 GATGTGCTCA CCATTACTCT GACTCCTAAG GTCACGTGTG TTGTGGTAGA 801CATCAGCAAG GATGATCCCG AGGTCCAGTT CAGCTGGTTT GTAGATGATG 851 TGGAGGTGCACACAGCTCAG ACGCAACCCC GGGAGGAGCA GTTCAACAGC 901 ACTTTCCGCT CAGTCAGTGAACTTCCCATC ATGCACCAGG ACTGGCTCAA 951 TGGCAAGGAG TTCAAATGCA GGGTCAACAGTGCAGCTTTC CCTGCCCCCA 1001 TCGAGAAAAC CATCTCCAAA ACCAAAGGCA GACCGAAGGCTCCACAGGTG 1051 TACACCATTC CACCTCCCAA GGAGCAGATG GCCAAGGATA AAGTCAGTCT1101 GACCTGCATG ATAACAGACT TCTTCCCTGA AGACATTACT GTGGAGTGGC 1151AGTGGAATGG GCAGCCAGCG GAGAACTACA AGAACACTCA GCCCATCATG 1201 GACACAGATGGCTCTTACTT CATCTACAGC AAGCTCAATG TGCAGAAGAG 1251 CAACTGGGAG GCAGGAAATACTTTCACCTG CTCTGTGTTA CATGAGGGCC 1301 TGCACAACCA CCATACTGAG AAGAGCCTCTCCCACTCTCC TGGTAAATGAAmino acid sequence of the Ab-3 HC including signal peptide:

(SEQ ID NO: 131) 1 MKCSWVIFFL MAVVTGVNSE VQLQQSGAEL VRPGALVKLSCTASDFNIKD 51 FYLHWMRQRP EQGLDWIGRI DPENGDTLYD PKFQDKATLT TDTSSNTAYL 101QLSGLTSETT AVYYCSREAD YFHDGTSYWY FDVWGAGTTI TVSSAKTTPP 151 SVYPLAPGSAAQTNSMVTLG CLVKGYFPEP VTVTWNSGSL SSGVHTFPAV 201 LQSDLYTLSS SVTVPSSTWPSETVTCNVAH PASSTKVDKK IVPRDCGCKP 251 CICTVPEVSS VFIFPPKPKD VLTITLTPKVTCVVVDISKD DPEVQFSWFV 301 DDVEVHTAQT QPREEQFNST FRSVSELPIM HQDWLNGKEFKCRVNSAAFP 351 APIEKTISKT KGRPKAPQVY TIPPPKEQMA KDKVSLTCMI TDFFPEDITV401 EWQWNGQPAE NYKNTQPIMD TDGSYFIYSK LNVQKSNWEA GNTFTCSVLH 451EGLHNHHTEK SLSHSPGKNucleic acid sequence of the Ab-3 HC including signal peptide encodingsequence:

(SEQ ID NO: 132) 1 ATGAAATGCA GCTGGGTCAT CTTCTTCCTG ATGGCAGTGGTTACAGGGGT 51 CAATTCAGAG GTTCAGCTGC AGCAGTCTGG GGCTGAACTT GTGAGGCCAG 101GGGCCTTAGT CAAGTTGTCC TGCACAGCTT CTGACTTCAA CATTAAAGAC 151 TTCTATCTACACTGGATGAG GCAGCGGCCT GAACAGGGCC TGGACTGGAT 201 TGGAAGGATT GATCCTGAGAATGGTGATAC TTTATATGAC CCGAAGTTCC 251 AGGACAAGGC CACTCTTACA ACAGACACATCCTCCAACAC AGCCTACCTG 301 CAGCTCAGCG GCCTGACATC TGAGACCACT GCCGTCTATTACTGTTCTAG 351 AGAGGCGGAT TATTTCCACG ATGGTACCTC CTACTGGTAC TTCGATGTCT401 GGGGCGCAGG GACCACAATC ACCGTCTCCT CAGCCAAAAC GACACCCCCA 451TCTGTCTATC CACTGGCCCC TGGATCTGCT GCCCAAACTA ACTCCATGGT 501 GACCCTGGGATGCCTGGTCA AGGGCTATTT CCCTGAGCCA GTGACAGTGA 551 CCTGGAACTC TGGATCCCTGTCCAGCGGTG TGCACACCTT CCCAGCTGTC 601 CTGCAGTCTG ACCTCTACAC TCTGAGCAGCTCAGTGACTG TCCCCTCCAG 651 CACCTGGCCC AGCGAGACCG TCACCTGCAA CGTTGCCCACCCGGCCAGCA 701 GCACCAAGGT GGACAAGAAA ATTGTGCCCA GGGATTGTGG TTGTAAGCCT751 TGCATATGTA CAGTCCCAGA AGTATCATCT GTCTTCATCT TCCCCCCAAA 801GCCCAAGGAT GTGCTCACCA TTACTCTGAC TCCTAAGGTC ACGTGTGTTG 851 TGGTAGACATCAGCAAGGAT GATCCCGAGG TCCAGTTCAG CTGGTTTGTA 901 GATGATGTGG AGGTGCACACAGCTCAGACG CAACCCCGGG AGGAGCAGTT 951 CAACAGCACT TTCCGCTCAG TCAGTGAACTTCCCATCATG CACCAGGACT 1001 GGCTCAATGG CAAGGAGTTC AAATGCAGGG TCAACAGTGCAGCTTTCCCT 1051 GCCCCCATCG AGAAAACCAT CTCCAAAACC AAAGGCAGAC CGAAGGCTCC1101 ACAGGTGTAC ACCATTCCAC CTCCCAAGGA GCAGATGGCC AAGGATAAAG 1151TCAGTCTGAC CTGCATGATA ACAGACTTCT TCCCTGAAGA CATTACTGTG 1201 GAGTGGCAGTGGAATGGGCA GCCAGCGGAG AACTACAAGA ACACTCAGCC 1251 CATCATGGAC ACAGATGGCTCTTACTTCAT CTACAGCAAG CTCAATGTGC 1301 AGAAGAGCAA CTGGGAGGCA GGAAATACTTTCACCTGCTC TGTGTTACAT 1351 GAGGGCCTGC ACAACCACCA TACTGAGAAG AGCCTCTCCCACTCTCCTGG 1401 TAAATGA

Ab-4

The sequences of the Antibody 4 (also referred to herein as Ab-4) LC andHC are as follows:

Ab-4 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-4 LC:

(SEQ ID NO: 133)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-4 LC:

(SEQ ID NO: 134) 1 GATATCCAGA TGACACAGAT TACATCCTCC CTGTCTGCCTCTCTGGGAGA 51 CAGGGTCTCC ATCAGTTGCA GGGCAAGTCA AGACATTAGC AATTATTTAA 101ACTGGTATCA GCAGAAACCA GATGGAACTT TTAAACTCCT TATCTTCTAC 151 ACATCAAGATTACTCTCAGG AGTCCCATCA AGGTTCAGTG GCAGTGGGTC 201 TGGAACAGAT TATTCTCTCACCATTTACAA CCTGGAGCAA GAAGATTTTG 251 CCACTTACTT TTGCCAACAG GGAGATACGCTTCCGTACAC TTTCGGAGGG 301 GGGACCAAGC TGGAAATAAA ACGGGCTGAT GCTGCACCAACTGTATCCAT 351 CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC TCAGTCGTGT401 GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA GTGGAAGATT 451GATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA CTGATCAGGA 501 CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG TTGACCAAGG 551 ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC TCACAAGACA 601 TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT GTTAGAmino acid sequence of the Ab-4 LC including signal peptide:

(SEQ ID NO: 135) 1 MMSSAQFLGL LLLCFQGTRC DIQMTQITSS LSASLGDRVSISCRASQDIS 51 NYLNWYQQKP DGTFKLLIFY TSRLLSGVPS RFSGSGSGTD YSLTIYNLEQ 101EDFATYFCQQ GDTLPYTFGG GTKLEIKRAD AAPTVSIFPP SSEQLTSGGA 151 SVVCFLNNFYPKDINVKWKI DGSERQNGVL NSWTDQDSKD STYSMSSTLT 201 LTKDEYERHN SYTCEATHKTSTSPIVKSFN RNECNucleic acid sequence of the Ab-4 LC including signal peptide encodingsequence:

(SEQ ID NO: 136) 1 ATGATGTCCT CTGCTCAGTT CCTTGGTCTC CTGTTGCTCTGTTTTCAAGG 51 TACCAGATGT GATATCCAGA TGACACAGAT TACATCCTCC CTGTCTGCCT 101CTCTGGGAGA CAGGGTCTCC ATCAGTTGCA GGGCAAGTCA AGACATTAGC 151 AATTATTTAAACTGGTATCA GCAGAAACCA GATGGAACTT TTAAACTCCT 201 TATCTTCTAC ACATCAAGATTACTCTCAGG AGTCCCATCA AGGTTCAGTG 251 GCAGTGGGTC TGGAACAGAT TATTCTCTCACCATTTACAA CCTGGAGCAA 301 GAAGATTTTG CCACTTACTT TTGCCAACAG GGAGATACGCTTCCGTACAC 351 TTTCGGAGGG GGGACCAAGC TGGAAATAAA ACGGGCTGAT GCTGCACCAA401 CTGTATCCAT CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC 451TCAGTCGTGT GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA 501 GTGGAAGATTGATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA 551 CTGATCAGGA CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG 601 TTGACCAAGG ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC 651 TCACAAGACA TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT 701 GTTAG

Ab-4 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-4 HC:

(SEQ ID NO: 137)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-4 HC:

(SEQ ID NO: 138) 1 GAGGTCCAAC TGCAACAGTC TGGACCTGAA CTAATGAAGCCTGGGGCTTC 51 AGTGAAGATG TCCTGCAAGG CTTCTGGATA TACATTCACT GACTACAACA 101TGCACTGGGT GAAGCAGAAC CAAGGAAAGA CCCTAGAGTG GATAGGAGAA 151 ATTAATCCTAACAGTGGTGG TGCTGGCTAC AACCAGAAGT TCAAGGGCAA 201 GGCCACATTG ACTGTAGACAAGTCCTCCAC CACAGCCTAC ATGGAGCTCC 251 GCAGCCTGAC ATCTGAGGAC TCTGCAGTCTATTACTGTGC AAGATTGGGC 301 TACGATGATA TCTACGACGA CTGGTACTTC GATGTCTGGGGCGCAGGGAC 351 CACGGTCACC GTCTCCTCAG CCAAAACGAC ACCCCCATCT GTCTATCCAC401 TGGCCCCTGG ATCTGCTGCC CAAACTAACT CCATGGTGAC CCTGGGATGC 451CTGGTCAAGG GCTATTTCCC TGAGCCAGTG ACAGTGACCT GGAACTCTGG 501 ATCCCTGTCCAGCGGTGTGC ACACCTTCCC AGCTGTCCTG CAGTCTGACC 551 TCTACACTCT GAGCAGCTCAGTGACTGTCC CCTCCAGCAC CTGGCCCAGC 601 GAGACCGTCA CCTGCAACGT TGCCCACCCGGCCAGCAGCA CCAAGGTGGA 651 CAAGAAAATT GTGCCCAGGG ATTGTGGTTG TAAGCCTTGCATATGTACAG 701 TCCCAGAAGT ATCATCTGTC TTCATCTTCC CCCCAAAGCC CAAGGATGTG751 CTCACCATTA CTCTGACTCC TAAGGTCACG TGTGTTGTGG TAGACATCAG 801CAAGGATGAT CCCGAGGTCC AGTTCAGCTG GTTTGTAGAT GATGTGGAGG 851 TGCACACAGCTCAGACGCAA CCCCGGGAGG AGCAGTTCAA CAGCACTTTC 901 CGCTCAGTCA GTGAACTTCCCATCATGCAC CAGGACTGGC TCAATGGCAA 951 GGAGTTCAAA TGCAGGGTCA ACAGTGCAGCTTTCCCTGCC CCCATCGAGA 1001 AAACCATCTC CAAAACCAAA GGCAGACCGA AGGCTCCACAGGTGTACACC 1051 ATTCCACCTC CCAAGGAGCA GATGGCCAAG GATAAAGTCA GTCTGACCTG1101 CATGATAACA GACTTCTTCC CTGAAGACAT TACTGTGGAG TGGCAGTGGA 1151ATGGGCAGCC AGCGGAGAAC TACAAGAACA CTCAGCCCAT CATGGACACA 1201 GATGGCTCTTACTTCATCTA CAGCAAGCTC AATGTGCAGA AGAGCAACTG 1251 GGAGGCAGGA AATACTTTCACCTGCTCTGT GTTACATGAG GGCCTGCACA 1301 ACCACCATAC TGAGAAGAGC CTCTCCCACTCTCCTGGTAA ATGAAmino acid sequence of the Ab-4 HC including signal peptide:

(SEQ ID NO: 139) 1MGWSWTFLFL LSGTAGVLSE VQLQQSGPEL MKPGASVKMS CKASGYTFTD 51YNMHWVKQNQ GKTLEWIGEI NPNSGGAGYN QKFKGKATLT VDKSSTTAYM 101ELRSLTSEDS AVYYCARLGY DDIYDDWYFD VWGAGTTVTV SSAKTTPPSV 151YPLAPGSAAQ TNSMVTLGCL VKGYFPEPVT VTWNSGSLSS GVHTFPAVLQ 201SDLYTLSSSV TVPSSTWPSE TVTCNVAHPA SSTKVDKKIV PRDCGCKPCI 251CTVPEVSSVF IFPPKPKDVL TITLTPKVTC VVVDISKDDP EVQFSWFVDD 301VEVHTAQTQP REEQFNSTFR SVSELPIMHQ DWLNGKEFKC RVNSAAFPAP 351IEKTISKTKG RPKAPQVYTI PPPKEQMAKD KVSLTCMITD FFPEDITVEW 401QWNGQPAENY KNTQPIMDTD GSYFIYSKLN VQKSNWEAGN TFTCSVLHEG 451LHNHHTEKSL SHSPGKNucleic acid sequence of the Ab-4 HC including signal peptide encodingsequence:

(SEQ ID NO: 140) 1ATGGGATGGA GCTGGACCTT TCTCTTCCTC CTGTCAGGAA CTGCAGGTGT 51CCTCTCTGAG GTCCAACTGC AACAGTCTGG ACCTGAACTA ATGAAGCCTG 101GGGCTTCAGT GAAGATGTCC TGCAAGGCTT CTGGATATAC ATTCACTGAC 151TACAACATGC ACTGGGTGAA GCAGAACCAA GGAAAGACCC TAGAGTGGAT 201AGGAGAAATT AATCCTAACA GTGGTGGTGC TGGCTACAAC CAGAAGTTCA 251AGGGCAAGGC CACATTGACT GTAGACAAGT CCTCCACCAC AGCCTACATG 301GAGCTCCGCA GCCTGACATC TGAGGACTCT GCAGTCTATT ACTGTGCAAG 351ATTGGGCTAC GATGATATCT ACGACGACTG GTACTTCGAT GTCTGGGGCG 401CAGGGACCAC GGTCACCGTC TCCTCAGCCA AAACGACACC CCCATCTGTC 451TATCCACTGG CCCCTGGATC TGCTGCCCAA ACTAACTCCA TGGTGACCCT 501GGGATGCCTG GTCAAGGGCT ATTTCCCTGA GCCAGTGACA GTGACCTGGA 551ACTCTGGATC CCTGTCCAGC GGTGTGCACA CCTTCCCAGC TGTCCTGCAG 601TCTGACCTCT ACACTCTGAG CAGCTCAGTG ACTGTCCCCT CCAGCACCTG 651GCCCAGCGAG ACCGTCACCT GCAACGTTGC CCACCCGGCC AGCAGCACCA 701AGGTGGACAA GAAAATTGTG CCCAGGGATT GTGGTTGTAA GCCTTGCATA 751TGTACAGTCC CAGAAGTATC ATCTGTCTTC ATCTTCCCCC CAAAGCCCAA 801GGATGTGCTC ACCATTACTC TGACTCCTAA GGTCACGTGT GTTGTGGTAG 851ACATCAGCAA GGATGATCCC GAGGTCCAGT TCAGCTGGTT TGTAGATGAT 901GTGGAGGTGC ACACAGCTCA GACGCAACCC CGGGAGGAGC AGTTCAACAG 951CACTTTCCGC TCAGTCAGTG AACTTCCCAT CATGCACCAG GACTGGCTCA 1001ATGGCAAGGA GTTCAAATGC AGGGTCAACA GTGCAGCTTT CCCTGCCCCC 1051ATCGAGAAAA CCATCTCCAA AACCAAAGGC AGACCGAAGG CTCCACAGGT 1101GTACACCATT CCACCTCCCA AGGAGCAGAT GGCCAAGGAT AAAGTCAGTC 1151TGACCTGCAT GATAACAGAC TTCTTCCCTG AAGACATTAC TGTGGAGTGG 1201CAGTGGAATG GGCAGCCAGC GGAGAACTAC AAGAACACTC AGCCCATCAT 1251GGACACAGAT GGCTCTTACT TCATCTACAG CAAGCTCAAT GTGCAGAAGA 1301GCAACTGGGA GGCAGGAAAT ACTTTCACCT GCTCTGTGTT ACATGAGGGC 1351CTGCACAACC ACCATACTGA GAAGAGCCTC TCCCACTCTC CTGGTAAATG 1401 AAb-4 was humanized to generate Ab-5.

Ab-5

The sequences of the Antibody 5 (also referred to herein as Ab-5) LC andHC are as follows:

Ab-5 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-5 LC:

(SEQ ID NO: 141)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-5 LC:

(SEQ ID NO: 142) 1GACATCCAGA TGACCCAGTC TCCATCCTCC CTCTCCGCAT CCGTAGGCGA 51CCGCGTAACC ATAACATGTA GAGCATCTCA AGATATTTCC AACTATTTGA 101ATTGGTACCA ACAAAAACCC GGCAAAGCAC CTAAACTCCT CATTTACTAT 151ACATCAAGAC TCCTCTCCGG CGTTCCATCA CGATTCTCAG GCTCCGGCTC 201CGGCACAGAT TTCACACTCA CTATTTCCTC CCTCCAACCA GAAGATTTTG 251CAACCTATTA CTGTCAACAA GGCGATACAC TCCCATACAC ATTCGGCGGC 301GGCACAAAAG TTGAAATTAA ACGTACGGTG GCTGCACCAT CTGTCTTCAT 351CTTCCCGCCA TCTGATGAGC AGTTGAAATC TGGAACTGCC TCTGTTGTGT 401GCCTGCTGAA TAACTTCTAT CCCAGAGAGG CCAAAGTACA GTGGAAGGTG 451GATAACGCCC TCCAATCGGG TAACTCCCAG GAGAGTGTCA CAGAGCAGGA 501CAGCAAGGAC AGCACCTACA GCCTCAGCAG CACCCTGACG CTGAGCAAAG 551CAGACTACGA GAAACACAAA GTCTACGCCT GCGAAGTCAC CCATCAGGGC 601CTGAGCTCGC CCGTCACAAA GAGCTTCAAC AGGGGAGAGT GTAmino acid sequence of the Ab-5 LC including signal peptide:

(SEQ ID NO: 143) 1MDMRVPAQLL GLLLLWLRGA RCDIQMTQSP SSLSASVGDR VTITCRASQD 51ISNYLNWYQQ KPGKAPKLLI YYTSRLLSGV PSRFSGSGSG TDFTLTISSL 101QPEDFATYYC QQGDTLPYTF GGGTKVEIKR TVAAPSVFIF PPSDEQLKSG 151TASVVCLLNN FYPREAKVQW KVDNALQSGN SQESVTEQDS KDSTYSLSST 201LTLSKADYEK HKVYACEVTH QGLSSPVTKS FNRGECNucleic acid sequence of the Ab-5 LC including signal peptide encodingsequence:

(SEQ ID NO: 144) 1ATGGACATGA GGGTCCCCGC TCAGCTCCTG GGGCTCCTGC TACTCTGGCT 51CCGAGGTGCC AGATGTGACA TCCAGATGAC CCAGTCTCCA TCCTCCCTCT 101CCGCATCCGT AGGCGACCGC GTAACCATAA CATGTAGAGC ATCTCAAGAT 151ATTTCCAACT ATTTGAATTG GTACCAACAA AAACCCGGCA AAGCACCTAA 201ACTCCTCATT TACTATACAT CAAGACTCCT CTCCGGCGTT CCATCACGAT 251TCTCAGGCTC CGGCTCCGGC ACAGATTTCA CACTCACTAT TTCCTCCCTC 301CAACCAGAAG ATTTTGCAAC CTATTACTGT CAACAAGGCG ATACACTCCC 351ATACACATTC GGCGGCGGCA CAAAAGTTGA AATTAAACGT ACGGTGGCTG 401CACCATCTGT CTTCATCTTC CCGCCATCTG ATGAGCAGTT GAAATCTGGA 451ACTGCCTCTG TTGTGTGCCT GCTGAATAAC TTCTATCCCA GAGAGGCCAA 501AGTACAGTGG AAGGTGGATA ACGCCCTCCA ATCGGGTAAC TCCCAGGAGA 551GTGTCACAGA GCAGGACAGC AAGGACAGCA CCTACAGCCT CAGCAGCACC 601CTGACGCTGA GCAAAGCAGA CTACGAGAAA CACAAAGTCT ACGCCTGCGA 651AGTCACCCAT CAGGGCCTGA GCTCGCCCGT CACAAAGAGC TTCAACAGGG 701 GAGAGTGT

Ab-5 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-5 HC:

(SEQ ID NO: 145)

Amino acid sequence of the mature form (signal peptide removed) of theAb-5 HC without carboxy-terminal lysine:

(SEQ ID NO: 392)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-5 HC:

(SEQ ID NO: 146) 1GAGGTGCAGC TGGTGCAGAG CGGCGCCGAG GTAAAAAAAC CAGGAGCAAG 51CGTTAAAGTT TCTTGTAAAG CAAGCGGATA TACATTTACA GATTACAACA 101TGCATTGGGT AAGACAAGCG CCAGGACAAG GATTGGAATG GATGGGCGAA 151ATTAACCCTA ATAGTGGAGG AGCAGGCTAC AATCAAAAAT TCAAAGGGAG 201AGTTACAATG ACAACAGACA CAAGCACTTC AACAGCATAT ATGGAACTGC 251GATCACTTAG AAGCGACGAT ACAGCTGTAT ACTATTGCGC ACGACTTGGG 301TATGATGATA TATATGATGA CTGGTATTTC GATGTTTGGG GCCAGGGAAC 351AACAGTTACC GTCTCTAGTG CCTCCACCAA GGGCCCATCG GTCTTCCCCC 401TGGCGCCCTG CTCCAGGAGC ACCTCCGAGA GCACAGCGGC CCTGGGCTGC 451CTGGTCAAGG ACTACTTCCC CGAACCGGTG ACGGTGTCGT GGAACTCAGG 501CGCTCTGACC AGCGGCGTGC ACACCTTCCC AGCTGTCCTA CAGTCCTCAG 551GACTCTACTC CCTCAGCAGC GTGGTGACCG TGCCCTCCAG CAACTTCGGC 601ACCCAGACCT ACACCTGCAA CGTAGATCAC AAGCCCAGCA ACACCAAGGT 651GGACAAGACA GTTGAGCGCA AATGTTGTGT CGAGTGCCCA CCGTGCCCAG 701CACCACCTGT GGCAGGACCG TCAGTCTTCC TCTTCCCCCC AAAACCCAAG 751GACACCCTCA TGATCTCCCG GACCCCTGAG GTCACGTGCG TGGTGGTGGA 801CGTGAGCCAC GAAGACCCCG AGGTCCAGTT CAACTGGTAC GTGGACGGCG 851TGGAGGTGCA TAATGCCAAG ACAAAGCCAC GGGAGGAGCA GTTCAACAGC 901ACGTTCCGTG TGGTCAGCGT CCTCACCGTT GTGCACCAGG ACTGGCTGAA 951CGGCAAGGAG TACAAGTGCA AGGTCTCCAA CAAAGGCCTC CCAGCCCCCA 1001TCGAGAAAAC CATCTCCAAA ACCAAAGGGC AGCCCCGAGA ACCACAGGTG 1051TACACCCTGC CCCCATCCCG GGAGGAGATG ACCAAGAACC AGGTCAGCCT 1101GACCTGCCTG GTCAAAGGCT TCTACCCCAG CGACATCGCC GTGGAGTGGG 1151AGAGCAATGG GCAGCCGGAG AACAACTACA AGACCACACC TCCCATGCTG 1201GACTCCGACG GCTCCTTCTT CCTCTACAGC AAGCTCACCG TGGACAAGAG 1251CAGGTGGCAG CAGGGGAACG TCTTCTCATG CTCCGTGATG CATGAGGCTC 1301TGCACAACCA CTACACGCAG AAGAGCCTCT CCCTGTCTCC GGGTAAAAmino acid sequence of the Ab-5 HC including signal peptide:

(SEQ ID NO: 147) 1MDWTWRILFL VAAATGAHSE VQLVQSGAEV KKPGASVKVS CKASGYTFTD 51YNMHWVRQAP GQGLEWMGEI NPNSGGAGYN QKFKGRVTMT TDTSTSTAYM 101ELRSLRSDDT AVYYCARLGY DDIYDDWYFD VWGQGTTVTV SSASTKGPSV 151FPLAPCSRST SESTAALGCL VKDYFPEPVT VSWNSGALTS GVHTFPAVLQ 201SSGLYSLSSV VTVPSSNFGT QTYTCNVDHK PSNTKVDKTV ERKCCVECPP 251CPAPPVAGPS VFLFPPKPKD TLMISRTPEV TCVVVDVSHE DPEVQFNWYV 301DGVEVHNAKT KPREEQFNST FRVVSVLTVV HQDWLNGKEY KCKVSNKGLP 351APIEKTISKT KGQPREPQVY TLPPSREEMT KNQVSLTCLV KGFYPSDIAV 401EWESNGQPEN NYKTTPPMLD SDGSFFLYSK LTVDKSRWQQ GNVFSCSVMH 451EALHNHYTQK SLSLSPGKNucleic acid sequence of the Ab-5 HC including signal peptide encodingsequence:

(SEQ ID NO: 148) 1ATGGACTGGA CCTGGAGGAT CCTCTTCTTG GTGGCAGCAG CCACAGGAGC 51CCACTCCGAG GTGCAGCTGG TGCAGAGCGG CGCCGAGGTA AAAAAACCAG 101GAGCAAGCGT TAAAGTTTCT TGTAAAGCAA GCGGATATAC ATTTACAGAT 151TACAACATGC ATTGGGTAAG ACAAGCGCCA GGACAAGGAT TGGAATGGAT 201GGGCGAAATT AACCCTAATA GTGGAGGAGC AGGCTACAAT CAAAAATTCA 251AAGGGAGAGT TACAATGACA ACAGACACAA GCACTTCAAC AGCATATATG 301GAACTGCGAT CACTTAGAAG CGACGATACA GCTGTATACT ATTGCGCACG 351ACTTGGGTAT GATGATATAT ATGATGACTG GTATTTCGAT GTTTGGGGCC 401AGGGAACAAC AGTTACCGTC TCTAGTGCCT CCACCAAGGG CCCATCGGTC 451TTCCCCCTGG CGCCCTGCTC CAGGAGCACC TCCGAGAGCA CAGCGGCCCT 501GGGCTGCCTG GTCAAGGACT ACTTCCCCGA ACCGGTGACG GTGTCGTGGA 551ACTCAGGCGC TCTGACCAGC GGCGTGCACA CCTTCCCAGC TGTCCTACAG 601TCCTCAGGAC TCTACTCCCT CAGCAGCGTG GTGACCGTGC CCTCCAGCAA 651CTTCGGCACC CAGACCTACA CCTGCAACGT AGATCACAAG CCCAGCAACA 701CCAAGGTGGA CAAGACAGTT GAGCGCAAAT GTTGTGTCGA GTGCCCACCG 751TGCCCAGCAC CACCTGTGGC AGGACCGTCA GTCTTCCTCT TCCCCCCAAA 801ACCCAAGGAC ACCCTCATGA TCTCCCGGAC CCCTGAGGTC ACGTGCGTGG 851TGGTGGACGT GAGCCACGAA GACCCCGAGG TCCAGTTCAA CTGGTACGTG 901GACGGCGTGG AGGTGCATAA TGCCAAGACA AAGCCACGGG AGGAGCAGTT 951CAACAGCACG TTCCGTGTGG TCAGCGTCCT CACCGTTGTG CACCAGGACT 1001GGCTGAACGG CAAGGAGTAC AAGTGCAAGG TCTCCAACAA AGGCCTCCCA 1051GCCCCCATCG AGAAAACCAT CTCCAAAACC AAAGGGCAGC CCCGAGAACC 1101ACAGGTGTAC ACCCTGCCCC CATCCCGGGA GGAGATGACC AAGAACCAGG 1151TCAGCCTGAC CTGCCTGGTC AAAGGCTTCT ACCCCAGCGA CATCGCCGTG 1201GAGTGGGAGA GCAATGGGCA GCCGGAGAAC AACTACAAGA CCACACCTCC 1251CATGCTGGAC TCCGACGGCT CCTTCTTCCT CTACAGCAAG CTCACCGTGG 1301ACAAGAGCAG GTGGCAGCAG GGGAACGTCT TCTCATGCTC CGTGATGCAT 1351GAGGCTCTGC ACAACCACTA CACGCAGAAG AGCCTCTCCC TGTCTCCGGG 1401 TAAA

Ab-5 Variable Domains:

Ab-5 light chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 376)

Ab-5 light chain variable domain DNA sequence (without signal sequence):

(SEQ ID NO: 377) 1GACATCCAGA TGACCCAGTC TCCATCCTCC CTCTCCGCAT CCGTAGGCGA 51CCGCGTAACC ATAACATGTA GAGCATCTCA AGATATTTCC AACTATTTGA 101ATTGGTACCA ACAAAAACCC GGCAAAGCAC CTAAACTCCT CATTTACTAT 151ACATCAAGAC TCCTCTCCGG CGTTCCATCA CGATTCTCAG GCTCCGGCTC 201CGGCACAGAT TTCACACTCA CTATTTCCTC CCTCCAACCA GAAGATTTTG 251CAACCTATTA CTGTCAACAA GGCGATACAC TCCCATACAC ATTCGGCGGC 301GGCACAAAAG TTGAAATTAA AAb-5 heavy chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 378)

Ab-5 heavy chain variable domain DNA sequence (without signal sequence):

(SEQ ID NO: 379) 1GAGGTGCAGC TGGTGCAGAG CGGCGCCGAG GTAAAAAAAC CAGGAGCAAG 51CGTTAAAGTT TCTTGTAAAG CAAGCGGATA TACATTTACA GATTACAACA 101TGCATTGGGT AAGACAAGCG CCAGGACAAG GATTGGAATG GATGGGCGAA 151ATTAACCCTA ATAGTGGAGG AGCAGGCTAC AATCAAAAAT TCAAAGGGAG 201AGTTACAATG ACAACAGACA CAAGCACTTC AACAGCATAT ATGGAACTGC 251GATCACTTAG AAGCGACGAT ACAGCTGTAT ACTATTGCGC ACGACTTGGG 301TATGATGATA TATATGATGA CTGGTATTTC GATGTTTGGG GCCAGGGAAC 351AACAGTTACC GTCTCTAGT

The CDR (complementarity determining region) sequences in the variableregion of the heavy chain of Ab-5 are as follows:

CDR-H1: (SEQ ID NO: 245) DYNMH CDR-H2: (SEQ ID NO: 246)EINPNSGGAGYNQKFKG CDR-H3: (SEQ ID NO: 247) LGYDDIYDDWYFDV

The light chain variable region CDR sequences of Ab-5 are:

CDR-L1: (SEQ ID NO: 78) RASQDISNYLN CDR-L2: (SEQ ID NO: 79) YTSRLLSCDR-L3: (SEQ ID NO: 80) QQGDTLPYT

Ab-6

The sequences of the Antibody 6 (also referred to herein as Ab-6) LC andHC are as follows:

Ab-6 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-6 LC:

(SEQ ID NO: 149)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-6 LC:

(SEQ ID NO: 150) 1GATATCCAGA TGACACAGAC TACATCCTCC CTGTCTGCCT CTCTGGGAGA 51CAGAGTCACC ATCAGTTGCA GGGCAAGTCA GGACATTAGC AATTATTTAA 101ACTGGTTTCA GCAGAAACCA GATGGAACTC TTAAACTCCT GATCTTCTAC 151ACATCAAGAT TACACTCAGG AGTTCCATCA AGGTTCAGTG GCAGTGGGTC 201TGGAACAGAT TATTCTCTCA CCATTAGCAA CCTGGAGCAA GAAGATATTG 251CCACTTACTT TTGCCAACAG GGTGATACGC TTCCGTACAC GTTCGGGGGG 301GGGACCAAGC TGGAAATAAG ACGGGCTGAT GCTGCACCAA CTGTATCCAT 351CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC TCAGTCGTGT 401GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA GTGGAAGATT 451GATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA CTGATCAGGA 501CAGCAAAGAC AGCACCTACA GCATGAGCAG CACCCTCACG TTGACCAAGG 551ACGAGTATGA ACGACATAAC AGCTATACCT GTGAGGCCAC TCACAAGACA 601TCAACTTCAC CCATTGTCAA GAGCTTCAAC AGGAATGAGT GTTAGAmino acid sequence of the Ab-6 LC including signal peptide:

(SEQ ID NO: 151) 1MMSSAQFLGL LLLCFQGTRC DIQMTQTTSS LSASLGDRVT ISCRASQDIS 51NYLNWFQQKP DGTLKLLIFY TSRLHSGVPS RFSGSGSGTD YSLTISNLEQ 101EDIATYFCQQ GDTLPYTFGG GTKLEIRRAD AAPTVSIFPP SSEQLTSGGA 151SVVCFLNNFY PKDINVKWKI DGSERQNGVL NSWTDQDSKD STYSMSSTLT 201LTKDEYERHN SYTCEATHKT STSPIVKSFN RNECNucleic acid sequence of the Ab-6 LC including signal peptide encodingsequence:

(SEQ ID NO: 152) 1ATGATGTCCT CTGCTCAGTT CCTTGGTCTC CTGTTGCTCT GTTTTCAAGG 51TACCAGATGT GATATCCAGA TGACACAGAC TACATCCTCC CTGTCTGCCT 101CTCTGGGAGA CAGAGTCACC ATCAGTTGCA GGGCAAGTCA GGACATTAGC 151AATTATTTAA ACTGGTTTCA GCAGAAACCA GATGGAACTC TTAAACTCCT 201GATCTTCTAC ACATCAAGAT TACACTCAGG AGTTCCATCA AGGTTCAGTG 251GCAGTGGGTC TGGAACAGAT TATTCTCTCA CCATTAGCAA CCTGGAGCAA 301GAAGATATTG CCACTTACTT TTGCCAACAG GGTGATACGC TTCCGTACAC 351GTTCGGGGGG GGGACCAAGC TGGAAATAAG ACGGGCTGAT GCTGCACCAA 401CTGTATCCAT CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC 451TCAGTCGTGT GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA 501GTGGAAGATT GATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA 551CTGATCAGGA CAGCAAAGAC AGCACCTACA GCATGAGCAG CACCCTCACG 601TTGACCAAGG ACGAGTATGA ACGACATAAC AGCTATACCT GTGAGGCCAC 651TCACAAGACA TCAACTTCAC CCATTGTCAA GAGCTTCAAC AGGAATGAGT 701 GTTAG

Ab-6 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-6 HC:

(SEQ ID NO: 153)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-6 HC:

(SEQ ID NO: 154) 1 GAGGTCCAGC TGCAACAGTC TGGACCTGAA CTAATGAAGCCTGGGGCTTC 51 AGTGAAGATG TCCTGCAAGG CTTCTGGATA CACATTCACT GACTACAACA 101TGCACTGGGT GAAACAGAAC CAAGGAAAGA GCCTAGAGTG GATAGGAGAA 151 ATTAATCCTAACAGTGGTGG TAGTGGCTAC AACCAAAAGT TCAAAGGCAA 201 GGCCACATTG ACTGTAGACAAGTCTTCCAG CACAGCCTAC ATGGAGCTCC 251 GCAGCCTGAC ATCTGAGGAC TCTGCAGTCTATTACTGTGC AAGATTGGTC 301 TACGATGGCA GCTACGAGGA CTGGTACTTC GATGTCTGGGGCGCAGGGAC 351 CACGGTCACC GTCTCCTCAG CCAAAACGAC ACCCCCATCT GTCTATCCAC401 TGGCCCCTGG ATCTGCTGCC CAAACTAACT CCATGGTGAC CCTGGGATGC 451CTGGTCAAGG GCTATTTCCC TGAGCCAGTG ACAGTGACCT GGAACTCTGG 501 ATCCCTGTCCAGCGGTGTGC ACACCTTCCC AGCTGTCCTG CAGTCTGACC 551 TCTACACTCT GAGCAGCTCAGTGACTGTCC CCTCCAGCAC CTGGCCCAGC 601 GAGACCGTCA CCTGCAACGT TGCCCACCCGGCCAGCAGCA CCAAGGTGGA 651 CAAGAAAATT GTGCCCAGGG ATTGTGGTTG TAAGCCTTGCATATGTACAG 701 TCCCAGAAGT ATCATCTGTC TTCATCTTCC CCCCAAAGCC CAAGGATGTG751 CTCACCATTA CTCTGACTCC TAAGGTCACG TGTGTTGTGG TAGACATCAG 801CAAGGATGAT CCCGAGGTCC AGTTCAGCTG GTTTGTAGAT GATGTGGAGG 851 TGCACACAGCTCAGACGCAA CCCCGGGAGG AGCAGTTCAA CAGCACTTTC 901 CGCTCAGTCA GTGAACTTCCCATCATGCAC CAGGACTGGC TCAATGGCAA 951 GGAGTTCAAA TGCAGGGTCA ACAGTGCAGCTTTCCCTGCC CCCATCGAGA 1001 AAACCATCTC CAAAACCAAA GGCAGACCGA AGGCTCCACAGGTGTACACC 1051 ATTCCACCTC CCAAGGAGCA GATGGCCAAG GATAAAGTCA GTCTGACCTG1101 CATGATAACA GACTTCTTCC CTGAAGACAT TACTGTGGAG TGGCAGTGGA 1151ATGGGCAGCC AGCGGAGAAC TACAAGAACA CTCAGCCCAT CATGGACACA 1201 GATGGCTCTTACTTCATCTA CAGCAAGCTC AATGTGCAGA AGAGCAACTG 1251 GGAGGCAGGA AATACTTTCACCTGCTCTGT GTTACATGAG GGCCTGCACA 1301 ACCACCATAC TGAGAAGAGC CTCTCCCACTCTCCTGGTAA ATGAAmino acid sequence of the Ab-6 HC including signal peptide:

(SEQ ID NO: 155) 1 MGWSWTFLFL LSGTAGVLSE VQLQQSGPEL MKPGASVKMSCKASGYTFTD 51 YNMHWVKQNQ GKSLEWIGEI NPNSGGSGYN QKFKGKATLT VDKSSSTAYM 101ELRSLTSEDS AVYYCARLVY DGSYEDWYFD VWGAGTTVTV SSAKTTPPSV 151 YPLAPGSAAQTNSMVTLGCL VKGYFPEPVT VTWNSGSLSS GVHTFPAVLQ 201 SDLYTLSSSV TVPSSTWPSETVTCNVAHPA SSTKVDKKIV PRDCGCKPCI 251 CTVPEVSSVF IFPPKPKDVL TITLTPKVTCVVVDISKDDP EVQFSWFVDD 301 VEVHTAQTQP REEQFNSTFR SVSELPIMHQ DWLNGKEFKCRVNSAAFPAP 351 IEKTISKTKG RPKAPQVYTI PPPKEQMAKD KVSLTCMITD FFPEDITVEW401 QWNGQPAENY KNTQPIMDTD GSYFIYSKLN VQKSNWEAGN TFTCSVLHEG 451LHNHHTEKSL SHSPGKNucleic acid sequence of the Ab-6 HC including signal peptide encodingsequence:

(SEQ ID NO: 156) 1 ATGGGATGGA GCTGGACCTT TCTCTTCCTC CTGTCAGGAACTGCAGGTGT 51 CCTCTCTGAG GTCCAGCTGC AACAGTCTGG ACCTGAACTA ATGAAGCCTG 101GGGCTTCAGT GAAGATGTCC TGCAAGGCTT CTGGATACAC ATTCACTGAC 151 TACAACATGCACTGGGTGAA ACAGAACCAA GGAAAGAGCC TAGAGTGGAT 201 AGGAGAAATT AATCCTAACAGTGGTGGTAG TGGCTACAAC CAAAAGTTCA 251 AAGGCAAGGC CACATTGACT GTAGACAAGTCTTCCAGCAC AGCCTACATG 301 GAGCTCCGCA GCCTGACATC TGAGGACTCT GCAGTCTATTACTGTGCAAG 351 ATTGGTCTAC GATGGCAGCT ACGAGGACTG GTACTTCGAT GTCTGGGGCG401 CAGGGACCAC GGTCACCGTC TCCTCAGCCA AAACGACACC CCCATCTGTC 451TATCCACTGG CCCCTGGATC TGCTGCCCAA ACTAACTCCA TGGTGACCCT 501 GGGATGCCTGGTCAAGGGCT ATTTCCCTGA GCCAGTGACA GTGACCTGGA 551 ACTCTGGATC CCTGTCCAGCGGTGTGCACA CCTTCCCAGC TGTCCTGCAG 601 TCTGACCTCT ACACTCTGAG CAGCTCAGTGACTGTCCCCT CCAGCACCTG 651 GCCCAGCGAG ACCGTCACCT GCAACGTTGC CCACCCGGCCAGCAGCACCA 701 AGGTGGACAA GAAAATTGTG CCCAGGGATT GTGGTTGTAA GCCTTGCATA751 TGTACAGTCC CAGAAGTATC ATCTGTCTTC ATCTTCCCCC CAAAGCCCAA 801GGATGTGCTC ACCATTACTC TGACTCCTAA GGTCACGTGT GTTGTGGTAG 851 ACATCAGCAAGGATGATCCC GAGGTCCAGT TCAGCTGGTT TGTAGATGAT 901 GTGGAGGTGC ACACAGCTCAGACGCAACCC CGGGAGGAGC AGTTCAACAG 951 CACTTTCCGC TCAGTCAGTG AACTTCCCATCATGCACCAG GACTGGCTCA 1001 ATGGCAAGGA GTTCAAATGC AGGGTCAACA GTGCAGCTTTCCCTGCCCCC 1051 ATCGAGAAAA CCATCTCCAA AACCAAAGGC AGACCGAAGG CTCCACAGGT1101 GTACACCATT CCACCTCCCA AGGAGCAGAT GGCCAAGGAT AAAGTCAGTC 1151TGACCTGCAT GATAACAGAC TTCTTCCCTG AAGACATTAC TGTGGAGTGG 1201 CAGTGGAATGGGCAGCCAGC GGAGAACTAC AAGAACACTC AGCCCATCAT 1251 GGACACAGAT GGCTCTTACTTCATCTACAG CAAGCTCAAT GTGCAGAAGA 1301 GCAACTGGGA GGCAGGAAAT ACTTTCACCTGCTCTGTGTT ACATGAGGGC 1351 CTGCACAACC ACCATACTGA GAAGAGCCTC TCCCACTCTCCTGGTAAATG 1401 A

Ab-7

The sequences of the Antibody 7 (also referred to herein as Ab-7) LC andHC are as follows:

Ab-7 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-7 LC:

(SEQ ID NO: 157)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-7 LC:

(SEQ ID NO: 158) 1 GATATCCAGA TGACACAGAC TACATCCTCC CTGTCTGCCTCTCTGGGAGA 51 CAGAGTCACC ATCTGTTGCA GGGCAAGTCA GGTCATTACC AATTATTTAT 101ACTGGTATCA GCAGAAACCA GATGGAACTT TTAAACTCCT GATCTACTAC 151 ACATCAAGATTACACTCAGG AGTCCCATCA AGGTTCAGTG GCAGTGGGTC 201 TGGAACAGAT TATTCTCTCACCATTAGCAA CCTGGAACAG GAAGATATTG 251 CCACTTACTT TTGCCAACAG GGTGATACGCTTCCGTACAC GTTCGGAGGG 301 GGGACCAAGC TGGAAATAAA ACGGGCTGAT GCTGCACCAACTGTATCCAT 351 CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC TCAGTCGTGT401 GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA GTGGAAGATT 451GATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA CTGATCAGGA 501 CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG TTGACCAAGG 551 ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC TCACAAGACA 601 TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT GTAmino acid sequence of the Ab-7 LC including signal peptide:

(SEQ ID NO: 159) 1 MMSSAQFLGL LLLCFQGTRC DIQMTQTTSS LSASLGDRVTICCRASQVIT 51 NYLYWYQQKP DGTFKLLIYY TSRLHSGVPS RFSGSGSGTD YSLTISNLEQ 101EDIATYFCQQ GDTLPYTFGG GTKLEIKRAD AAPTVSIFPP SSEQLTSGGA 151 SVVCFLNNFYPKDINVKWKI DGSERQNGVL NSWTDQDSKD STYSMSSTLT 201 LTKDEYERHN SYTCEATHKTSTSPIVKSFN RNECNucleic acid sequence of the Ab-7 LC including signal peptide encodingsequence:

(SEQ ID NO: 160) 1 ATGATGTCCT CTGCTCAGTT CCTTGGTCTC CTGTTGCTCTGTTTTCAAGG 51 TACCAGATGT GATATCCAGA TGACACAGAC TACATCCTCC CTGTCTGCCT 101CTCTGGGAGA CAGAGTCACC ATCTGTTGCA GGGCAAGTCA GGTCATTACC 151 AATTATTTATACTGGTATCA GCAGAAACCA GATGGAACTT TTAAACTCCT 201 GATCTACTAC ACATCAAGATTACACTCAGG AGTCCCATCA AGGTTCAGTG 251 GCAGTGGGTC TGGAACAGAT TATTCTCTCACCATTAGCAA CCTGGAACAG 301 GAAGATATTG CCACTTACTT TTGCCAACAG GGTGATACGCTTCCGTACAC 351 GTTCGGAGGG GGGACCAAGC TGGAAATAAA ACGGGCTGAT GCTGCACCAA401 CTGTATCCAT CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC 451TCAGTCGTGT GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA 501 GTGGAAGATTGATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA 551 CTGATCAGGA CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG 601 TTGACCAAGG ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC 651 TCACAAGACA TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT 701 GT

Ab-7 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-7 HC:

(SEQ ID NO: 161)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-7 HC:

(SEQ ID NO: 162) 1 GAGGTCCAGC TGCAACAGTC TGGACCTGAA CTAATGAAGCCTGGGGCTTC 51 AGTGAAGATG TCCTGCAAGG CTTCTGGATA CACATTCACT GACTACAACA 101TGCACTGGAT GAAGCAGAAC CAAGGAAAGA GCCTAGAATG GATAGGAGAA 151 ATTAATCCTAACAGTGGTGG TGCTGGCTAC AACCAGCAGT TCAAAGGCAA 201 GGCCACATTG ACTGTAGACAAGTCCTCCAG GACAGCCTAC ATGGAGCTCC 251 GCAGCCTGAC ATCTGAGGAC TCTGCAGTCTATTACTGTGC AAGATTGGGC 301 TACGTTGGTA ATTACGAGGA CTGGTACTTC GATGTCTGGGGCGCAGGGAC 351 CACGGTCACC GTCTCCTCAG CCAAAACGAC ACCCCCATCT GTCTATCCAC401 TGGCCCCTGG ATCTGCTGCC CAAACTAACT CCATGGTGAC CCTGGGATGC 451CTGGTCAAGG GCTATTTCCC TGAGCCAGTG ACAGTGACCT GGAACTCTGG 501 ATCCCTGTCCAGCGGTGTGC ACACCTTCCC AGCTGTCCTG CAGTCTGACC 551 TCTACACTCT GAGCAGCTCAGTGACTGTCC CCTCCAGCAC CTGGCCCAGC 601 GAGACCGTCA CCTGCAACGT TGCCCACCCGGCCAGCAGCA CCAAGGTGGA 651 CAAGAAAATT GTGCCCAGGG ATTGTGGTTG TAAGCCTTGCATATGTACAG 701 TCCCAGAAGT ATCATCTGTC TTCATCTTCC CCCCAAAGCC CAAGGATGTG751 CTCACCATTA CTCTGACTCC TAAGGTCACG TGTGTTGTGG TAGACATCAG 801CAAGGATGAT CCCGAGGTCC AGTTCAGCTG GTTTGTAGAT GATGTGGAGG 851 TGCACACAGCTCAGACGCAA CCCCGGGAGG AGCAGTTCAA CAGCACTTTC 901 CGCTCAGTCA GTGAACTTCCCATCATGCAC CAGGACTGGC TCAATGGCAA 951 GGAGTTCAAA TGCAGGGTCA ACAGTGCAGCTTTCCCTGCC CCCATCGAGA 1001 AAACCATCTC CAAAACCAAA GGCAGACCGA AGGCTCCACAGGTGTACACC 1051 ATTCCACCTC CCAAGGAGCA GATGGCCAAG GATAAAGTCA GTCTGACCTG1101 CATGATAACA GACTTCTTCC CTGAAGACAT TACTGTGGAG TGGCAGTGGA 1151ATGGGCAGCC AGCGGAGAAC TACAAGAACA CTCAGCCCAT CATGGACACA 1201 GATGGCTCTTACTTCATCTA CAGCAAGCTC AATGTGCAGA AGAGCAACTG 1251 GGAGGCAGGA AATACTTTCACCTGCTCTGT GTTACATGAG GGCCTGCACA 1301 ACCACCATAC TGAGAAGAGC CTCTCCCACTCTCCTGGTAA AAmino acid sequence of the Ab-7 HC including signal peptide:

(SEQ ID NO: 163) 1 MGWSWTFLFL LSGTAGVLSE VQLQQSGPEL MKPGASVKMSCKASGYTFTD 51 YNMHWMKQNQ GKSLEWIGEI NPNSGGAGYN QQFKGKATLT VDKSSRTAYM 101ELRSLTSEDS AVYYCARLGY VGNYEDWYFD VWGAGTTVTV SSAKTTPPSV 151 YPLAPGSAAQTNSMVTLGCL VKGYFPEPVT VTWNSGSLSS GVHTFPAVLQ 201 SDLYTLSSSV TVPSSTWPSETVTCNVAHPA SSTKVDKKIV PRDCGCKPCI 251 CTVPEVSSVF IFPPKPKDVL TITLTPKVTCVVVDISKDDP EVQFSWFVDD 301 VEVHTAQTQP REEQFNSTFR SVSELPIMHQ DWLNGKEFKCRVNSAAFPAP 351 IEKTISKTKG RPKAPQVYTI PPPKEQMAKD KVSLTCMITD FFPEDITVEW401 QWNGQPAENY KNTQPIMDTD GSYFIYSKLN VQKSNWEAGN TFTCSVLHEG 451LHNHHTEKSL SHSPGKNucleic acid sequence of the Ab-7 HC including signal peptide encodingsequence:

(SEQ ID NO: 164) 1 ATGGGATGGA GCTGGACCTT TCTCTTCCTC CTGTCAGGAACTGCAGGTGT 51 CCTCTCTGAG GTCCAGCTGC AACAGTCTGG ACCTGAACTA ATGAAGCCTG 101GGGCTTCAGT GAAGATGTCC TGCAAGGCTT CTGGATACAC ATTCACTGAC 151 TACAACATGCACTGGATGAA GCAGAACCAA GGAAAGAGCC TAGAATGGAT 201 AGGAGAAATT AATCCTAACAGTGGTGGTGC TGGCTACAAC CAGCAGTTCA 251 AAGGCAAGGC CACATTGACT GTAGACAAGTCCTCCAGGAC AGCCTACATG 301 GAGCTCCGCA GCCTGACATC TGAGGACTCT GCAGTCTATTACTGTGCAAG 351 ATTGGGCTAC GTTGGTAATT ACGAGGACTG GTACTTCGAT GTCTGGGGCG401 CAGGGACCAC GGTCACCGTC TCCTCAGCCA AAACGACACC CCCATCTGTC 451TATCCACTGG CCCCTGGATC TGCTGCCCAA ACTAACTCCA TGGTGACCCT 501 GGGATGCCTGGTCAAGGGCT ATTTCCCTGA GCCAGTGACA GTGACCTGGA 551 ACTCTGGATC CCTGTCCAGCGGTGTGCACA CCTTCCCAGC TGTCCTGCAG 601 TCTGACCTCT ACACTCTGAG CAGCTCAGTGACTGTCCCCT CCAGCACCTG 651 GCCCAGCGAG ACCGTCACCT GCAACGTTGC CCACCCGGCCAGCAGCACCA 701 AGGTGGACAA GAAAATTGTG CCCAGGGATT GTGGTTGTAA GCCTTGCATA751 TGTACAGTCC CAGAAGTATC ATCTGTCTTC ATCTTCCCCC CAAAGCCCAA 801GGATGTGCTC ACCATTACTC TGACTCCTAA GGTCACGTGT GTTGTGGTAG 851 ACATCAGCAAGGATGATCCC GAGGTCCAGT TCAGCTGGTT TGTAGATGAT 901 GTGGAGGTGC ACACAGCTCAGACGCAACCC CGGGAGGAGC AGTTCAACAG 951 CACTTTCCGC TCAGTCAGTG AACTTCCCATCATGCACCAG GACTGGCTCA 1001 ATGGCAAGGA GTTCAAATGC AGGGTCAACA GTGCAGCTTTCCCTGCCCCC 1051 ATCGAGAAAA CCATCTCCAA AACCAAAGGC AGACCGAAGG CTCCACAGGT1101 GTACACCATT CCACCTCCCA AGGAGCAGAT GGCCAAGGAT AAAGTCAGTC 1151TGACCTGCAT GATAACAGAC TTCTTCCCTG AAGACATTAC TGTGGAGTGG 1201 CAGTGGAATGGGCAGCCAGC GGAGAACTAC AAGAACACTC AGCCCATCAT 1251 GGACACAGAT GGCTCTTACTTCATCTACAG CAAGCTCAAT GTGCAGAAGA 1301 GCAACTGGGA GGCAGGAAAT ACTTTCACCTGCTCTGTGTT ACATGAGGGC 1351 CTGCACAACC ACCATACTGA GAAGAGCCTC TCCCACTCTCCTGGTAAA

Ab-8

The sequences of the Antibody 8 (also referred to herein as Ab-8) LC andHC are as follows:

Ab-8 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-8

(SEQ ID NO: 165)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-8 LC:

(SEQ ID NO: 166) 1 GATATCCAGA TGACACAGAC TACATCCTCC CTGTCTGCCTCTCTGGGAGA 51 CAGGGTCTCC ATCAGTTGCA GGGCAAGTCA AGACATTAGC AATTATTTAA 101ACTGGTATCA GCAGAAACCA GATGGAACTT TTAAACTCCT TATCTTCTAC 151 ACATCAAGATTACTCTCAGG AGTCCCATCA AGGTTCAGTG GCAGTGGGTC 201 TGGAACAGAT TATTCTCTCACCATTTACAA CCTGGAGCAA GAAGATTTTG 251 CCACTTACTT TTGCCAACAG GGAGATACGCTTCCGTACAC TTTCGGAGGG 301 GGGACCAAAC TGGAAATAAA ACGGGCTGAT GCTGCACCAACTGTATCCAT 351 CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC TCAGTCGTGT401 GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA GTGGAAGATT 451GATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA CTGATCAGGA 501 CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG TTGACCAAGG 551 ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC TCACAAGACA 601 TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT GTTAGAmino acid sequence of the Ab-8 LC including signal peptide:

(SEQ ID NO: 167) 1 MMSSAQFLGL LLLCFQGTRC DIQMTQTTSS LSASLGDRVSISCRASQDIS 51 NYLNWYQQKP DGTFKLLIFY TSRLLSGVPS RFSGSGSGTD YSLTIYNLEQ 101EDFATYFCQQ GDTLPYTFGG GTKLEIKRAD AAPTVSIFPP SSEQLTSGGA 151 SVVCFLNNFYPKDINVKWKI DGSERQNGVL NSWTDQDSKD STYSMSSTLT 201 LTKDEYERHN SYTCEATHKTSTSPIVKSFN RNECNucleic acid sequence of the Ab-8 LC including signal peptide encodingsequence:

(SEQ ID NO: 168) 1 ATGATGTCCT CTGCTCAGTT CCTTGGTCTC CTGTTGCTCTGTTTTCAAGG 51 TACCAGATGT GATATCCAGA TGACACAGAC TACATCCTCC CTGTCTGCCT 101CTCTGGGAGA CAGGGTCTCC ATCAGTTGCA GGGCAAGTCA AGACATTAGC 151 AATTATTTAAACTGGTATCA GCAGAAACCA GATGGAACTT TTAAACTCCT 201 TATCTTCTAC ACATCAAGATTACTCTCAGG AGTCCCATCA AGGTTCAGTG 251 GCAGTGGGTC TGGAACAGAT TATTCTCTCACCATTTACAA CCTGGAGCAA 301 GAAGATTTTG CCACTTACTT TTGCCAACAG GGAGATACGCTTCCGTACAC 351 TTTCGGAGGG GGGACCAAAC TGGAAATAAA ACGGGCTGAT GCTGCACCAA401 CTGTATCCAT CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC 451TCAGTCGTGT GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA 501 GTGGAAGATTGATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA 551 CTGATCAGGA CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG 601 TTGACCAAGG ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC 651 TCACAAGACA TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT 701 GTTAG

Ab-8 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-8 HC:

(SEQ ID NO: 169)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-8 HC:

(SEQ ID NO: 170) 1 GAGGTCCAAC TGCAACAGTC TGGACCTGAA CTAATGAAGCCTGGGGCTTC 51 AGTGAAGATG TCCTGCAAGG CTTCTGGATA TACATTCACT GACTACAACA 101TGCACTGGGT GAAGCAGAAC CAAGGAAAGA CCCTAGACTG GATAGGAGAA 151 ATTAATCCTAACAGTGGTGG TGCTGGCTAC AACCAGAAGT TCAAGGGCAA 201 GGCCACATTG ACTGTAGACAAGTCCTCCAC CACAGCCTAC ATGGAGCTCC 251 GCAGCCTGAC ATCTGAGGAC TCTGCAGTCTATTACTGTGC AAGATTGGGC 301 TACGATGATA TCTACGACGA CTGGTACTTC GATGTCTGGGGCGCAGGGAC 351 CACGGTCACC GTCTCCTCAG CCAAAACGAC ACCCCCATCT GTCTATCCAC401 TGGCCCCTGG ATCTGCTGCC CAAACTAACT CCATGGTGAC CCTGGGATGC 451CTGGTCAAGG GCTATTTCCC TGAGCCAGTG ACAGTGACCT GGAACTCTGG 501 ATCCCTGTCCAGCGGTGTGC ACACCTTCCC AGCTGTCCTG CAGTCTGACC 551 TCTACACTCT GAGCAGCTCAGTGACTGTCC CCTCCAGCAC CTGGCCCAGC 601 GAGACCGTCA CCTGCAACGT TGCCCACCCGGCCAGCAGCA CCAAGGTGGA 651 CAAGAAAATT GTGCCCAGGG ATTGTGGTTG TAAGCCTTGCATATGTACAG 701 TCCCAGAAGT ATCATCTGTC TTCATCTTCC CCCCAAAGCC CAAGGATGTG751 CTCACCATTA CTCTGACTCC TAAGGTCACG TGTGTTGTGG TAGACATCAG 801CAAGGATGAT CCCGAGGTCC AGTTCAGCTG GTTTGTAGAT GATGTGGAGG 851 TGCACACAGCTCAGACGCAA CCCCGGGAGG AGCAGTTCAA CAGCACTTTC 901 CGCTCAGTCA GTGAACTTCCCATCATGCAC CAGGACTGGC TCAATGGCAA 951 GGAGTTCAAA TGCAGGGTCA ACAGTGCAGCTTTCCCTGCC CCCATCGAGA 1001 AAACCATCTC CAAAACCAAA GGCAGACCGA AGGCTCCACAGGTGTACACC 1051 ATTCCACCTC CCAAGGAGCA GATGGCCAAG GATAAAGTCA GTCTGACCTG1101 CATGATAACA GACTTCTTCC CTGAAGACAT TACTGTGGAG TGGCAGTGGA 1151ATGGGCAGCC AGCGGAGAAC TACAAGAACA CTCAGCCCAT CATGGACACA 1201 GATGGCTCTTACTTCATCTA CAGCAAGCTC AATGTGCAGA AGAGCAACTG 1251 GGAGGCAGGA AATACTTTCACCTGCTCTGT GTTACATGAG GGCCTGCACA 1301 ACCACCATAC TGAGAAGAGC CTCTCCCACTCTCCTGGTAA ATGAAmino acid sequence of the Ab-8 HC including signal peptide:

(SEQ ID NO: 171) 1 MGWSWTFLFL LSGTAGVLSE VQLQQSGPEL MKPGASVKMSCKASGYTFTD 51 YNMHWVKQNQ GKTLDWIGEI NPNSGGAGYN QKFKGKATLT VDKSSTTAYM 101ELRSLTSEDS AVYYCARLGY DDIYDDWYFD VWGAGTTVTV SSAKTTPPSV 151 YPLAPGSAAQTNSMVTLGCL VKGYFPEPVT VTWNSGSLSS GVHTFPAVLQ 201 SDLYTLSSSV TVPSSTWPSETVTCNVAHPA SSTKVDKKIV PRDCGCKPCI 251 CTVPEVSSVF IFPPKPKDVL TITLTPKVTCVVVDISKDDP EVQFSWFVDD 301 VEVHTAQTQP REEQFNSTFR SVSELPIMHQ DWLNGKEFKCRVNSAAFPAP 351 IEKTISKTKG RPKAPQVYTI PPPKEQMAKD KVSLTCMITD FFPEDITVEW401 QWNGQPAENY KNTQPIMDTD GSYFIYSKLN VQKSNWEAGN TFTCSVLHEG 451LHNHHTEKSL SHSPGKNucleic acid sequence of the Ab-8 HC including signal peptide encodingsequence:

(SEQ ID NO: 172) 1 ATGGGATGGA GCTGGACCTT TCTCTTCCTC CTGTCAGGAACTGCAGGTGT 51 CCTCTCTGAG GTCCAACTGC AACAGTCTGG ACCTGAACTA ATGAAGCCTG 101GGGCTTCAGT GAAGATGTCC TGCAAGGCTT CTGGATATAC ATTCACTGAC 151 TACAACATGCACTGGGTGAA GCAGAACCAA GGAAAGACCC TAGACTGGAT 201 AGGAGAAATT AATCCTAACAGTGGTGGTGC TGGCTACAAC CAGAAGTTCA 251 AGGGCAAGGC CACATTGACT GTAGACAAGTCCTCCACCAC AGCCTACATG 301 GAGCTCCGCA GCCTGACATC TGAGGACTCT GCAGTCTATTACTGTGCAAG 351 ATTGGGCTAC GATGATATCT ACGACGACTG GTACTTCGAT GTCTGGGGCG401 CAGGGACCAC GGTCACCGTC TCCTCAGCCA AAACGACACC CCCATCTGTC 451TATCCACTGG CCCCTGGATC TGCTGCCCAA ACTAACTCCA TGGTGACCCT 501 GGGATGCCTGGTCAAGGGCT ATTTCCCTGA GCCAGTGACA GTGACCTGGA 551 ACTCTGGATC CCTGTCCAGCGGTGTGCACA CCTTCCCAGC TGTCCTGCAG 601 TCTGACCTCT ACACTCTGAG CAGCTCAGTGACTGTCCCCT CCAGCACCTG 651 GCCCAGCGAG ACCGTCACCT GCAACGTTGC CCACCCGGCCAGCAGCACCA 701 AGGTGGACAA GAAAATTGTG CCCAGGGATT GTGGTTGTAA GCCTTGCATA751 TGTACAGTCC CAGAAGTATC ATCTGTCTTC ATCTTCCCCC CAAAGCCCAA 801GGATGTGCTC ACCATTACTC TGACTCCTAA GGTCACGTGT GTTGTGGTAG 851 ACATCAGCAAGGATGATCCC GAGGTCCAGT TCAGCTGGTT TGTAGATGAT 901 GTGGAGGTGC ACACAGCTCAGACGCAACCC CGGGAGGAGC AGTTCAACAG 951 CACTTTCCGC TCAGTCAGTG AACTTCCCATCATGCACCAG GACTGGCTCA 1001 ATGGCAAGGA GTTCAAATGC AGGGTCAACA GTGCAGCTTTCCCTGCCCCC 1051 ATCGAGAAAA CCATCTCCAA AACCAAAGGC AGACCGAAGG CTCCACAGGT1101 GTACACCATT CCACCTCCCA AGGAGCAGAT GGCCAAGGAT AAAGTCAGTC 1151TGACCTGCAT GATAACAGAC TTCTTCCCTG AAGACATTAC TGTGGAGTGG 1201 CAGTGGAATGGGCAGCCAGC GGAGAACTAC AAGAACACTC AGCCCATCAT 1251 GGACACAGAT GGCTCTTACTTCATCTACAG CAAGCTCAAT GTGCAGAAGA 1301 GCAACTGGGA GGCAGGAAAT ACTTTCACCTGCTCTGTGTT ACATGAGGGC 1351 CTGCACAACC ACCATACTGA GAAGAGCCTC TCCCACTCTCCTGGTAAATG 1401 A

Ab-9

The sequences of the Antibody 9 (also referred to herein as Ab-9) LC andHC are as follows:

Ab-9 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-9 LC:

(SEQ ID NO: 173)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-9 LC:

(SEQ ID NO: 174) 1 GATATCCAGA TGACACAGAT TACATCCTCC CTGTCTGCCTCTCTGGGAGA 51 CAGGGTCTCC ATCAGTTGCA GGGCAAGTCA AGACATTAGC AATTATTTAA 101ATTGGTATCA GCAGAAACCA GATGGAACTT TTAAACTCCT TATCTTCTAC 151 ACATCAAGATTATTTTCAGG AGTCCCATCA AGGTTCAGTG GCAGTGGGTC 201 TGGAACAGAT TATTCTCTCACCATTTACAA CCTGGAGCAA GAAGATTTTG 251 CCACTTACTT TTGCCAACAG GGAGATACGCTTCCGTACAC TTTCGGAGGG 301 GGGACCAAGG TGGAAATAAA ACGGGCTGAT GCTGCACCAACTGTATCCAT 351 CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC TCAGTCGTGT401 GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA GTGGAAGATT 451GATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA CTGATCAGGA 501 CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG TTGACCAAGG 551 ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC TCACAAGACA 601 TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT GTAmino acid sequence of the Ab-9 LC including signal peptide:

(SEQ ID NO: 175) 1 MMSSAQFLGL LLLCFQGTRC DIQMTQITSS LSASLGDRVSISCRASQDIS 51 NYLNWYQQKP DGTFKLLIFY TSRLFSGVPS RFSGSGSGTD YSLTIYNLEQ 101EDFATYFCQQ GDTLPYTFGG GTKVEIKRAD AAPTVSIFPP SSEQLTSGGA 151 SVVCFLNNFYPKDINVKWKI DGSERQNGVL NSWTDQDSKD STYSMSSTLT 201 LTKDEYERHN SYTCEATHKTSTSPIVKSFN RNECNucleic acid sequence of the Ab-9 LC including signal peptide encodingsequence:

(SEQ ID NO: 176) 1 ATGATGTCCT CTGCTCAGTT CCTTGGTCTC CTGTTGCTCTGTTTTCAAGG 51 TACCAGATGT GATATCCAGA TGACACAGAT TACATCCTCC CTGTCTGCCT 101CTCTGGGAGA CAGGGTCTCC ATCAGTTGCA GGGCAAGTCA AGACATTAGC 151 AATTATTTAAATTGGTATCA GCAGAAACCA GATGGAACTT TTAAACTCCT 201 TATCTTCTAC ACATCAAGATTATTTTCAGG AGTCCCATCA AGGTTCAGTG 251 GCAGTGGGTC TGGAACAGAT TATTCTCTCACCATTTACAA CCTGGAGCAA 301 GAAGATTTTG CCACTTACTT TTGCCAACAG GGAGATACGCTTCCGTACAC 351 TTTCGGAGGG GGGACCAAGG TGGAAATAAA ACGGGCTGAT GCTGCACCAA401 CTGTATCCAT CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC 451TCAGTCGTGT GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA 501 GTGGAAGATTGATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA 551 CTGATCAGGA CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG 601 TTGACCAAGG ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC 651 TCACAAGACA TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT 701 GT

Ab-9 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-9 HC.

(SEQ ID NO: 177)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-9 HC:

(SEQ ID NO: 178) 1 GAGGTCCAAC TGCAACAGTC TGGACCTGAA CTAATGAAGCCTGGGACTTC 51 AGTGAAGATG TCCTGCAAGG CTTCTGGATA TACATTCACT GACTACAACA 101TGCACTGGGT GAAGCAGACC CAAGGAAAGA CCCTAGAGTG GATAGGAGAA 151 ATTAATCCTAACAGTGGTGG TGCTGGCTAC AACCAGAAGT TCAAGGGCAA 201 GGCCACATTG ACTGTAGACAAGTCCTCCAC CACAGCCTAC ATGGAGCTCC 251 GCAGCCTGAC ATCTGAGGAC TCTGCAGTCTATTACTGTGC AAAATTGGGC 301 TACGATGATA TCTACGACGA CTGGTATTTC GATGTCTGGGGCGCAGGGAC 351 CACGGTCACC GTCTCCTCAG CCAAAACAAC AGCCCCATCG GTCTATCCAC401 TGGCCCCTGT GTGTGGAGAT ACAACTGGCT CCTCGGTGAC TCTAGGATGC 451CTGGTCAAGG GTTATTTCCC TGAGCCAGTG ACCTTGACCT GGAACTCTGG 501 ATCCCTGTCCAGTGATGTGC ACACCTTCCC AGCTCTCCTG CAGTCTGGCC 551 TCTACACCCT CAGCAGCTCAGTGACTGTAA CCACCTGGCC CAGCCAGACC 601 ATCACCTGCA ATGTGGCCCA CCCGGCAAGCAGCACCAAAG TGGACAAGAA 651 AATTGAGCCC AGAGGGTCCC CAACACATAA ACCCTGTCCTCCATGCCCAG 701 CTCCTAACCT CTTGGGTGGA CCATCCGTCT TCATCTTCCC TCCAAAGATC751 AAGGATGTAC TCATGATCTC CCTGAGCCCC ATGGTCACGT GTGTGGTGGT 801GGATGTGAGC GAGGATGACC CAGATGTCCA TGTCAGCTGG TTCGTGAACA 851 ACGTGGAAGTACACACAGCT CAGACACAAA CCCATAGAGA GGATTACAAC 901 AGTACTATCC GGGTGGTCAGTGCCCTCCCC ATCCAGCACC AGGACTGGAT 951 GAGTGGCAAG GAGTTCAAAT GCAAGGTCAACAACAAAGCC CTCCCAGCGC 1001 CCATCGAGAG AACCATCTCA AAACCCAAAG GGCCAGTAAGAGCTCCACAG 1051 GTATATGTCT TGCCTCCACC AGAAGAAGAG ATGACTAAGA AACAGGTCAC1101 TCTGACCTGC ATGATCACAG ACTTCATGCC TGAAGACATT TACGTGGAGT 1151GGACCAACAA CGGGCAAACA GAGCTAAACT ACAAGAACAC TGAACCAGTC 1201 CTGGACTCTGATGGTTCTTA CTTCATGTAC AGCAAGCTGA GAGTGGAAAA 1251 GAAGAACTGG GTGGAAAGAAATAGCTACTC CTGTTCAGTG GTCCACGAGG 1301 GTCTGCACAA TCACCACACG ACTAAGAGCTTCTCCCGGAC TCCGGGTAAAAmino acid sequence of the Ab-9 HC including signal peptide:

(SEQ ID NO: 179) 1 MGWSWTFLFL LSGTAGVLSE VQLQQSGPEL MKPGTSVKMSCKASGYTFTD 51 YNMHWVKQTQ GKTLEWIGEI NPNSGGAGYN QKFKGKATLT VDKSSTTAYM 101ELRSLTSEDS AVYYCAKLGY DDIYDDWYFD VWGAGTTVTV SSAKTTAPSV 151 YPLAPVCGDTTGSSVTLGCL VKGYFPEPVT LTWNSGSLSS DVHTFPALLQ 201 SGLYTLSSSV TVTTWPSQTITCNVAHPASS TKVDKKIEPR GSPTHKPCPP 251 CPAPNLLGGP SVFIFPPKIK DVLMISLSPMVTCVVVDVSE DDPDVHVSWF 301 VNNVEVHTAQ TQTHREDYNS TIRVVSALPI QHQDWMSGKEFKCKVNNKAL 351 PAPIERTISK PKGPVRAPQV YVLPPPEEEM TKKQVTLTCM ITDFMPEDIY401 VEWTNNGQTE LNYKNTEPVL DSDGSYFMYS KLRVEKKNWV ERNSYSCSVV 451HEGLHNHHTT KSFSRTPGKNucleic acid sequence of the Ab-9 HC including signal peptide encodingsequence:

(SEQ ID NO: 180) 1 ATGGGATGGA GCTGGACCTT TCTCTTCCTC CTGTCAGGAACTGCAGGTGT 51 CCTCTCTGAG GTCCAACTGC AACAGTCTGG ACCTGAACTA ATGAAGCCTG 101GGACTTCAGT GAAGATGTCC TGCAAGGCTT CTGGATATAC ATTCACTGAC 151 TACAACATGCACTGGGTGAA GCAGACCCAA GGAAAGACCC TAGAGTGGAT 201 AGGAGAAATT AATCCTAACAGTGGTGGTGC TGGCTACAAC CAGAAGTTCA 251 AGGGCAAGGC CACATTGACT GTAGACAAGTCCTCCACCAC AGCCTACATG 301 GAGCTCCGCA GCCTGACATC TGAGGACTCT GCAGTCTATTACTGTGCAAA 351 ATTGGGCTAC GATGATATCT ACGACGACTG GTATTTCGAT GTCTGGGGCG401 CAGGGACCAC GGTCACCGTC TCCTCAGCCA AAACAACAGC CCCATCGGTC 451TATCCACTGG CCCCTGTGTG TGGAGATACA ACTGGCTCCT CGGTGACTCT 501 AGGATGCCTGGTCAAGGGTT ATTTCCCTGA GCCAGTGACC TTGACCTGGA 551 ACTCTGGATC CCTGTCCAGTGATGTGCACA CCTTCCCAGC TCTCCTGCAG 601 TCTGGCCTCT ACACCCTCAG CAGCTCAGTGACTGTAACCA CCTGGCCCAG 651 CCAGACCATC ACCTGCAATG TGGCCCACCC GGCAAGCAGCACCAAAGTGG 701 ACAAGAAAAT TGAGCCCAGA GGGTCCCCAA CACATAAACC CTGTCCTCCA751 TGCCCAGCTC CTAACCTCTT GGGTGGACCA TCCGTCTTCA TCTTCCCTCC 801AAAGATCAAG GATGTACTCA TGATCTCCCT GAGCCCCATG GTCACGTGTG 851 TGGTGGTGGATGTGAGCGAG GATGACCCAG ATGTCCATGT CAGCTGGTTC 901 GTGAACAACG TGGAAGTACACACAGCTCAG ACACAAACCC ATAGAGAGGA 951 TTACAACAGT ACTATCCGGG TGGTCAGTGCCCTCCCCATC CAGCACCAGG 1001 ACTGGATGAG TGGCAAGGAG TTCAAATGCA AGGTCAACAACAAAGCCCTC 1051 CCAGCGCCCA TCGAGAGAAC CATCTCAAAA CCCAAAGGGC CAGTAAGAGC1101 TCCACAGGTA TATGTCTTGC CTCCACCAGA AGAAGAGATG ACTAAGAAAC 1151AGGTCACTCT GACCTGCATG ATCACAGACT TCATGCCTGA AGACATTTAC 1201 GTGGAGTGGACCAACAACGG GCAAACAGAG CTAAACTACA AGAACACTGA 1251 ACCAGTCCTG GACTCTGATGGTTCTTACTT CATGTACAGC AAGCTGAGAG 1301 TGGAAAAGAA GAACTGGGTG GAAAGAAATAGCTACTCCTG TTCAGTGGTC 1351 CACGAGGGTC TGCACAATCA CCACACGACT AAGAGCTTCTCCCGGACTCC 1401 GGGTAAA

Ab-10

The sequences of the Antibody 10 (also referred to herein as Ab-10) LCand HC are as follows:

Ab-10 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-10 LC:

(SEQ ID NO: 181)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-10 LC:

(SEQ ID NO: 182) 1 GATATCCAGA TGACACAGAC TACATCCTCC CTGTCTGCCTCTCTGGGAGA 51 CAGGGTCTCC ATCAGTTGCA GGGCAAGTCA AGACATTAGC AATTATTTAA 101ACTGGTATCA GCAGAAACCA GATGGAACTT TTAAACTCCT TATCTTCTAC 151 ACATCAAGATTACTCTCAGG AGTCCCATCA AGGTTCAGTG GCAGTGGGTC 201 TGGAACAGAT TATTCTCTCACCATTTACAA CCTGGAGCAA GAAGATTTTG 251 CCACTTACTT TTGCCAACAG GGAGATACGCTTCCGTACAC TTTCGGAGGG 301 GGGACCAAAC TGGAAATAAA ACGGGCTGAT GCTGCACCAACTGTATCCAT 351 CTTCCCACTA TCCAGTGAGC AGTTAACATC TGGAGGTGCC TCAGTCGTGT401 GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA GTGGAAGATT 451GATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA CTGATCAGGA 501 CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG TTGACCAAGG 551 ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC TCACAAGACA 601 TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT GTTAGAmino acid sequence of the Ab-10 LC including signal peptide:

(SEQ ID NO: 183) 1 MMSSAQFLGL LLLCFQGTRC DIQMTQTTSS LSASLGDRVSISCRASQDIS 51 NYLNWYQQKP DGTFKLLIFY TSRLLSGVPS RFSGSGSGTD YSLTIYNLEQ 101EDFATYFCQQ GDTLPYTFGG GTKLEIKRAD AAPTVSIFPL SSEQLTSGGA 151 SVVCFLNNFYPKDINVKWKI DGSERQNGVL NSWTDQDSKD STYSMSSTLT 201 LTKDEYERHN SYTCEATHKTSTSPIVKSFN RNECNucleic acid sequence of the Ab-10 LC including signal peptide encodingsequence:

(SEQ ID NO: 184) 1 ATGATGTCCT CTGCTCAGTT CCTTGGTCTC CTGTTGCTCTGTTTTCAAGG 51 TACCAGATGT GATATCCAGA TGACACAGAC TACATCCTCC CTGTCTGCCT 101CTCTGGGAGA CAGGGTCTCC ATCAGTTGCA GGGCAAGTCA AGACATTAGC 151 AATTATTTAAACTGGTATCA GCAGAAACCA GATGGAACTT TTAAACTCCT 201 TATCTTCTAC ACATCAAGATTACTCTCAGG AGTCCCATCA AGGTTCAGTG 251 GCAGTGGGTC TGGAACAGAT TATTCTCTCACCATTTACAA CCTGGAGCAA 301 GAAGATTTTG CCACTTACTT TTGCCAACAG GGAGATACGCTTCCGTACAC 351 TTTCGGAGGG GGGACCAAAC TGGAAATAAA ACGGGCTGAT GCTGCACCAA401 CTGTATCCAT CTTCCCACTA TCCAGTGAGC AGTTAACATC TGGAGGTGCC 451TCAGTCGTGT GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA 501 GTGGAAGATTGATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA 551 CTGATCAGGA CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG 601 TTGACCAAGG ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC 651 TCACAAGACA TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT 701 GTTAG

Ab-10 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-10 HC:

(SEQ ID NO: 185)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-10 HC:

(SEQ ID NO: 186) 1 GAGGTCCAAC TGCAACAGTC TGGACCTGAA CTAATGAAGCCTGGGGCTTC 51 AGTGAAGATG TCCTGCAAGG CTTCTGGATA TACATTCACT GACTACAACA 101TGCACTGGGT GAAGCAGAAC CAAGGAAAGA CCCTAGAATG GATAGGAGAA 151 ATTAATCCTAACAGTGGTGG TGCTGGCTAC AACCAGAAGT TCAAGGGCAA 201 GGCCACATTG ACTGTAGACAAGTCCTCCAC CACAGCCTAC ATGGAGCTCC 251 GCAGCCTGAC ATCTGAGGAC TCTGCAGTCTATTACTGTGC AAGATTGGGC 301 TACGATGATA TCTACGACGA CTGGTACTTC GATGTCTGGGGCGCAGGGAC 351 CACGGTCACC GTCTCCTCAG CCAAAACGAC ACCCCCATCT GTCTATCCAC401 TGGCCCCTGG ATCTGCTGCC CAAACTAACT CCATGGTGAC CCTGGGATGC 451CTGGTCAAGG GCTATTTCCC TGAGCCAGTG ACAGTGACCT GGAACTCTGG 501 ATCCCTGTCCAGCGGTGTGC ACACCTTCCC AGCTGTCCTG CAGTCTGACC 551 TCTACACTCT GAGCAGCTCAGTGACTGTCC CCTCCAGCAC CTGGCCCAGC 601 GAGACCGTCA CCTGCAACGT TGCCCACCCGGCCAGCAGCA CCAAGGTGGA 651 CAAGAAAATT GTGCCCAGGG ATTGTGGTTG TAAGCCTTGCATATGTACAG 701 TCCCAGAAGT ATCATCTGTC TTCATCTTCC CCCCAAAGCC CAAGGATGTG751 CTCACCATTA CTCTGACTCC TAAGGTCACG TGTGTTGTGG TAGACATCAG 801CAAGGATGAT CCCGAGGTCC AGTTCAGCTG GTTTGTAGAT GATGTGGAGG 851 TGCACACAGCTCAGACGCAA CCCCGGGAGG AGCAGTTCAA CAGCACTTTC 901 CGCTCAGTCA GTGAACTTCCCATCATGCAC CAGGACTGGC TCAATGGCAA 951 GGAGTTCAAA TGCAGGGTCA ACAGTGCAGCTTTCCCTGCC CCCATCGAGA 1001 AAACCATCTC CAAAACCAAA GGCAGACCGA AGGCTCCACAGGTGTACACC 1051 ATTCCACCTC CCAAGGAGCA GATGGCCAAG GATAAAGTCA GTCTGACCTG1101 CATGATAACA GACTTCTTCC CTGAAGACAT TACTGTGGAG TGGCAGTGGA 1151ATGGGCAGCC AGCGGAGAAC TACAAGAACA CTCAGCCCAT CATGGACACA 1201 GATGGCTCTTACTTCATCTA CAGCAAGCTC AATGTGCAGA AGAGCAACTG 1251 GGAGGCAGGA AATACTTTCACCTGCTCTGT GTTACATGAG GGCCTGCACA 1301 ACCACCATAC TGAGAAGAGC CTCTCCCACTCTCCTGGTAA ATGAAmino acid sequence of the Ab-10 HC including signal peptide:

(SEQ ID NO: 187) 1 MGWSWTFLFL LSGTAGVLSE VQLQQSGPEL MKPGASVKMSCKASGYTFTD 51 YNMHWVKQNQ GKTLEWIGEI NPNSGGAGYN QKFKGKATLT VDKSSTTAYM 101ELRSLTSEDS AVYYCARLGY DDIYDDWYFD VWGAGTTVTV SSAKTTPPSV 151 YPLAPGSAAQTNSMVTLGCL VKGYFPEPVT VTWNSGSLSS GVHTFPAVLQ 201 SDLYTLSSSV TVPSSTWPSETVTCNVAHPA SSTKVDKKIV PRDCGCKPCI 251 CTVPEVSSVF IFPPKPKDVL TITLTPKVTCVVVDISKDDP EVQFSWFVDD 301 VEVHTAQTQP REEQFNSTFR SVSELPIMHQ DWLNGKEFKCRVNSAAFPAP 351 IEKTISKTKG RPKAPQVYTI PPPKEQMAKD KVSLTCMITD FFPEDITVEW401 QWNGQPAENY KNTQPIMDTD GSYFIYSKLN VQKSNWEAGN TFTCSVLHEG 451LHNHHTEKSL SHSPGKNucleic acid sequence of the Ab-10 HC including signal peptide encodingsequence:

(SEQ ID NO: 188) 1 ATGGGATGGA GCTGGACCTT TCTCTTCCTC CTGTCAGGAACTGCAGGTGT 51 CCTCTCTGAG GTCCAACTGC AACAGTCTGG ACCTGAACTA ATGAAGCCTG 101GGGCTTCAGT GAAGATGTCC TGCAAGGCTT CTGGATATAC ATTCACTGAC 151 TACAACATGCACTGGGTGAA GCAGAACCAA GGAAAGACCC TAGAATGGAT 201 AGGAGAAATT AATCCTAACAGTGGTGGTGC TGGCTACAAC CAGAAGTTCA 251 AGGGCAAGGC CACATTGACT GTAGACAAGTCCTCCACCAC AGCCTACATG 301 GAGCTCCGCA GCCTGACATC TGAGGACTCT GCAGTCTATTACTGTGCAAG 351 ATTGGGCTAC GATGATATCT ACGACGACTG GTACTTCGAT GTCTGGGGCG401 CAGGGACCAC GGTCACCGTC TCCTCAGCCA AAACGACACC CCCATCTGTC 451TATCCACTGG CCCCTGGATC TGCTGCCCAA ACTAACTCCA TGGTGACCCT 501 GGGATGCCTGGTCAAGGGCT ATTTCCCTGA GCCAGTGACA GTGACCTGGA 551 ACTCTGGATC CCTGTCCAGCGGTGTGCACA CCTTCCCAGC TGTCCTGCAG 601 TCTGACCTCT ACACTCTGAG CAGCTCAGTGACTGTCCCCT CCAGCACCTG 651 GCCCAGCGAG ACCGTCACCT GCAACGTTGC CCACCCGGCCAGCAGCACCA 701 AGGTGGACAA GAAAATTGTG CCCAGGGATT GTGGTTGTAA GCCTTGCATA751 TGTACAGTCC CAGAAGTATC ATCTGTCTTC ATCTTCCCCC CAAAGCCCAA 801GGATGTGCTC ACCATTACTC TGACTCCTAA GGTCACGTGT GTTGTGGTAG 851 ACATCAGCAAGGATGATCCC GAGGTCCAGT TCAGCTGGTT TGTAGATGAT 901 GTGGAGGTGC ACACAGCTCAGACGCAACCC CGGGAGGAGC AGTTCAACAG 951 CACTTTCCGC TCAGTCAGTG AACTTCCCATCATGCACCAG GACTGGCTCA 1001 ATGGCAAGGA GTTCAAATGC AGGGTCAACA GTGCAGCTTTCCCTGCCCCC 1051 ATCGAGAAAA CCATCTCCAA AACCAAAGGC AGACCGAAGG CTCCACAGGT1101 GTACACCATT CCACCTCCCA AGGAGCAGAT GGCCAAGGAT AAAGTCAGTC 1151TGACCTGCAT GATAACAGAC TTCTTCCCTG AAGACATTAC TGTGGAGTGG 1201 CAGTGGAATGGGCAGCCAGC GGAGAACTAC AAGAACACTC AGCCCATCAT 1251 GGACACAGAT GGCTCTTACTTCATCTACAG CAAGCTCAAT GTGCAGAAGA 1301 GCAACTGGGA GGCAGGAAAT ACTTTCACCTGCTCTGTGTT ACATGAGGGC 1351 CTGCACAACC ACCATACTGA GAAGAGCCTC TCCCACTCTCCTGGTAAATG 1401 A

Ab-11

The sequences of the Antibody 11 (also referred to herein as Ab-11) LCand HC are as follows:

Ab-11 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-11 LC:

(SEQ ID NO: 189)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-11 LC:

(SEQ ID NO: 190) 1 CAAATTGTTC TCTCCCAGTC TCCAGCATTC CTGTCTGTATCTCCAGGGGA 51 TAAGGTCACA ATGACTTGCA GGGCCAGCTC AAGTATAAGT TACATACACT 101GGTTTCAGCA GAAGCCAGGA TCCTCCCCCA GATCCTGGAT TTATGCCACA 151 TCCAACCTGGCTTCTGGAGT CCCTGGTCGC TTCAGTGGCA GTGGGTCTGG 201 GACCTCTTAC TCTCTCACAATCAGCAGAGT GGAGGCTGAG GATGCTGCCA 251 CTTATTACTG CCAGCAGTGG AGTAGTGACCCACTCACGTT CGGTGCTGGG 301 ACCAAGCTGG AGCTGAAACG GGCTGATGCT GCACCAACTGTATCCATCTT 351 CCCACCATCC AGTGAGCAGT TAACATCTGG AGGTGCCTCA GTCGTGTGCT401 TCTTGAACAA CTTCTACCCC AAAGACATCA ATGTCAAGTG GAAGATTGAT 451GGCAGTGAAC GACAAAATGG CGTCCTGAAC AGTTGGACTG ATCAGGACAG 501 CAAAGACAGCACCTACAGCA TGAGCAGCAC CCTCACGTTG ACCAAGGACG 551 AGTATGAACG ACATAACAGCTATACCTGTG AGGCCACTCA CAAGACATCA 601 ACTTCACCCA TTGTCAAGAG CTTCAACAGGAATGAGTGTT AGAmino acid sequence of the Ab-11 LC including signal peptide:

(SEQ ID NO: 191) 1 MDFQVQIFSF LLISASVIMS RGQIVLSQSP AFLSVSPGDKVTMTCRASSS 51 ISYIHWFQQK PGSSPRSWIY ATSNLASGVP GRFSGSGSGT SYSLTISRVE 101AEDAATYYCQ QWSSDPLTFG AGTKLELKRA DAAPTVSIFP PSSEQLTSGG 151 ASVVCFLNNFYPKDINVKWK IDGSERQNGV LNSWTDQDSK DSTYSMSSTL 201 TLTKDEYERH NSYTCEATHKTSTSPIVKSF NRNECNucleic acid sequence of the Ab-11 LC including signal peptide encodingsequence:

(SEQ ID NO: 192) 1 ATGGATTTTC AAGTGCAGAT TTTCAGCTTC CTGCTAATCAGTGCTTCAGT 51 CATAATGTCC AGAGGACAAA TTGTTCTCTC CCAGTCTCCA GCATTCCTGT 101CTGTATCTCC AGGGGATAAG GTCACAATGA CTTGCAGGGC CAGCTCAAGT 151 ATAAGTTACATACACTGGTT TCAGCAGAAG CCAGGATCCT CCCCCAGATC 201 CTGGATTTAT GCCACATCCAACCTGGCTTC TGGAGTCCCT GGTCGCTTCA 251 GTGGCAGTGG GTCTGGGACC TCTTACTCTCTCACAATCAG CAGAGTGGAG 301 GCTGAGGATG CTGCCACTTA TTACTGCCAG CAGTGGAGTAGTGACCCACT 351 CACGTTCGGT GCTGGGACCA AGCTGGAGCT GAAACGGGCT GATGCTGCAC401 CAACTGTATC CATCTTCCCA CCATCCAGTG AGCAGTTAAC ATCTGGAGGT 451GCCTCAGTCG TGTGCTTCTT GAACAACTTC TACCCCAAAG ACATCAATGT 501 CAAGTGGAAGATTGATGGCA GTGAACGACA AAATGGCGTC CTGAACAGTT 551 GGACTGATCA GGACAGCAAAGACAGCACCT ACAGCATGAG CAGCACCCTC 601 ACGTTGACCA AGGACGAGTA TGAACGACATAACAGCTATA CCTGTGAGGC 651 CACTCACAAG ACATCAACTT CACCCATTGT CAAGAGCTTCAACAGGAATG 701 AGTGTTAG

Ab-11 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-11 HC:

(SEQ ID NO: 193)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-11 HC:

(SEQ ID NO: 194) 1 GAAGTTCAGC TGCAACAGTC TGGGGCAGAC CTTGTGCAGCCAGGGGCCTC 51 AGTCAAGGTG TCCTGCACAG CTTCTGGCTT CGACATTAAG GACTACTATA 101TACACTGGAT GAAACAGAGG CCTGACCAGG GCCTGGAGTG GATTGGAAGG 151 GTTGATCCTGACAATGGTGA GACTGAATTT GCCCCGAAGT TCCCGGGCAA 201 GGCCACTTTT ACAACAGACACATCCTCCAA CACAGCCTAC CTACAACTCA 251 GAGGCCTGAC ATCTGAGGAC ACTGCCATCTATTACTGTGG GAGAGAAGAC 301 TACGATGGTA CCTACACCTG GTTTCCTTAT TGGGGCCAAGGGACTCTGGT 351 CACTGTCTCT GCAGCCAAAA CGACACCCCC ATCTGTCTAT CCACTGGCCC401 CTGGATCTGC TGCCCAAACT AACTCCATGG TGACCCTGGG ATGCCTGGTC 451AAGGGCTATT TCCCTGAGCC AGTGACAGTG ACCTGGAACT CTGGATCCCT 501 GTCCAGCGGTGTGCACACCT TCCCAGCTGT CCTGCAGTCT GACCTCTACA 551 CTCTGAGCAG CTCAGTGACTGTCCCCTCCA GCACCTGGCC CAGCGAGACC 601 GTCACCTGCA ACGTTGCCCA CCCGGCCAGCAGCACCAAGG TGGACAAGAA 651 AATTGTGCCC AGGGATTGTG GTTGTAAGCC TTGCATATGTACAGTCCCAG 701 AAGTATCATC TGTCTTCATC TTCCCCCCAA AGCCCAAGGA TGTGCTCACC751 ATTACTCTGA CTCCTAAGGT CACGTGTGTT GTGGTAGACA TCAGCAAGGA 801TGATCCCGAG GTCCAGTTCA GCTGGTTTGT AGATGATGTG GAGGTGCACA 851 CAGCTCAGACGCAACCCCGG GAGGAGCAGT TCAACAGCAC TTTCCGCTCA 901 GTCAGTGAAC TTCCCATCATGCACCAGGAC TGGCTCAATG GCAAGGAGTT 951 CAAATGCAGG GTCAACAGTG CAGCTTTCCCTGCCCCCATC GAGAAAACCA 1001 TCTCCAAAAC CAAAGGCAGA CCGAAGGCTC CACAGGTGTACACCATTCCA 1051 CCTCCCAAGG AGCAGATGGC CAAGGATAAA GTCAGTCTGA CCTGCATGAT1101 AACAGACTTC TTCCCTGAAG ACATTACTGT GGAGTGGCAG TGGAATGGGC 1151AGCCAGCGGA GAACTACAAG AACACTCAGC CCATCATGGA CACAGATGGC 1201 TCTTACTTCATCTACAGCAA GCTCAATGTG CAGAAGAGCA ACTGGGAGGC 1251 AGGAAATACT TTCACCTGCTCTGTGTTACA TGAGGGCCTG CACAACCACC 1301 ATACTGAGAA GAGCCTCTCC CACTCTCCTGGTAAATGAAmino acid sequence of the Ab-11 HC including signal peptide:

(SEQ ID NO: 195) 1 MKCSWVIFFL MAVVTGVNSE VQLQQSGADL VQPGASVKVSCTASGFDIKD 51 YYIHWMKQRP DQGLEWIGRV DPDNGETEFA PKFPGKATFT TDTSSNTAYL 101QLRGLTSEDT AIYYCGREDY DGTYTWFPYW GQGTLVTVSA AKTTPPSVYP 151 LAPGSAAQTNSMVTLGCLVK GYFPEPVTVT WNSGSLSSGV HTFPAVLQSD 201 LYTLSSSVTV PSSTWPSETVTCNVAHPASS TKVDKKIVPR DCGCKPCICT 251 VPEVSSVFIF PPKPKDVLTI TLTPKVTCVVVDISKDDPEV QFSWFVDDVE 301 VHTAQTQPRE EQFNSTFRSV SELPIMHQDW LNGKEFKCRVNSAAFPAPIE 351 KTISKTKGRP KAPQVYTIPP PKEQMAKDKV SLTCMITDFF PEDITVEWQW401 NGQPAENYKN TQPIMDTDGS YFIYSKLNVQ KSNWEAGNTF TCSVLHEGLH 451NHHTEKSLSH SPGKNucleic acid sequence of the Ab-11 HC including signal peptide encodingsequence:

(SEQ ID NO: 196) 1 ATGAAATGCA GCTGGGTCAT CTTCTTCCTG ATGGCAGTGGTTACAGGGGT 51 CAATTCAGAA GTTCAGCTGC AACAGTCTGG GGCAGACCTT GTGCAGCCAG 101GGGCCTCAGT CAAGGTGTCC TGCACAGCTT CTGGCTTCGA CATTAAGGAC 151 TACTATATACACTGGATGAA ACAGAGGCCT GACCAGGGCC TGGAGTGGAT 201 TGGAAGGGTT GATCCTGACAATGGTGAGAC TGAATTTGCC CCGAAGTTCC 251 CGGGCAAGGC CACTTTTACA ACAGACACATCCTCCAACAC AGCCTACCTA 301 CAACTCAGAG GCCTGACATC TGAGGACACT GCCATCTATTACTGTGGGAG 351 AGAAGACTAC GATGGTACCT ACACCTGGTT TCCTTATTGG GGCCAAGGGA401 CTCTGGTCAC TGTCTCTGCA GCCAAAACGA CACCCCCATC TGTCTATCCA 451CTGGCCCCTG GATCTGCTGC CCAAACTAAC TCCATGGTGA CCCTGGGATG 501 CCTGGTCAAGGGCTATTTCC CTGAGCCAGT GACAGTGACC TGGAACTCTG 551 GATCCCTGTC CAGCGGTGTGCACACCTTCC CAGCTGTCCT GCAGTCTGAC 601 CTCTACACTC TGAGCAGCTC AGTGACTGTCCCCTCCAGCA CCTGGCCCAG 651 CGAGACCGTC ACCTGCAACG TTGCCCACCC GGCCAGCAGCACCAAGGTGG 701 ACAAGAAAAT TGTGCCCAGG GATTGTGGTT GTAAGCCTTG CATATGTACA751 GTCCCAGAAG TATCATCTGT CTTCATCTTC CCCCCAAAGC CCAAGGATGT 801GCTCACCATT ACTCTGACTC CTAAGGTCAC GTGTGTTGTG GTAGACATCA 851 GCAAGGATGATCCCGAGGTC CAGTTCAGCT GGTTTGTAGA TGATGTGGAG 901 GTGCACACAG CTCAGACGCAACCCCGGGAG GAGCAGTTCA ACAGCACTTT 951 CCGCTCAGTC AGTGAACTTC CCATCATGCACCAGGACTGG CTCAATGGCA 1001 AGGAGTTCAA ATGCAGGGTC AACAGTGCAG CTTTCCCTGCCCCCATCGAG 1051 AAAACCATCT CCAAAACCAA AGGCAGACCG AAGGCTCCAC AGGTGTACAC1101 CATTCCACCT CCCAAGGAGC AGATGGCCAA GGATAAAGTC AGTCTGACCT 1151GCATGATAAC AGACTTCTTC CCTGAAGACA TTACTGTGGA GTGGCAGTGG 1201 AATGGGCAGCCAGCGGAGAA CTACAAGAAC ACTCAGCCCA TCATGGACAC 1251 AGATGGCTCT TACTTCATCTACAGCAAGCT CAATGTGCAG AAGAGCAACT 1301 GGGAGGCAGG AAATACTTTC ACCTGCTCTGTGTTACATGA GGGCCTGCAC 1351 AACCACCATA CTGAGAAGAG CCTCTCCCAC TCTCCTGGTAAATGA

Ab-12

The sequences of the Antibody 12 (also referred to herein as Ab-12) LCand HC are as follows:

Ab-12 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-12 LC:

(SEQ ID NO: 197)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-12 LC:

(SEQ ID NO: 198) 1 GATCTCCAGA TGACACAGAC TACTTCCTCC CTGTCTGCCTCTCTGGGAGA 51 CAGAGTCACC ATCAGTTGCA GGGCAAGTCA GGACATTAGC AATTATTTAA 101ACTGGTATCA GCAGAAACCA GATGGAACTG TTAAGCTCCT GATCTTCTAC 151 ACATCAACATTACAGTCAGG AGTCCCATCG AGGTTCAGTG GCAGTGGGTC 201 TGGAACAAAT TATTCTCTCACCATTACCAA CCTGGAGCAA GATGATGCTG 251 CCACTTACTT TTGCCAACAG GGTGATACGCTTCCGTACAC GTTCGGAGGG 301 GGGACCAAGC TGGAAATAAA ACGGGCTGAT GCTGCACCAACTGTATCCAT 351 CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC TCAGTCGTGT401 GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA GTGGAAGATT 451GATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA CTGATCAGGA 501 CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG TTGACCAAGG 551 ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC TCACAAGACA 601 TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT GTTAGAmino acid sequence of the Ab-12 LC including signal peptide:

(SEQ ID NO: 199) 1 MMSSAQFLGL LLLCFQGSRC DLQMTQTTSS LSASLGDRVTISCRASQDIS 51 NYLNWYQQKP DGTVKLLIFY TSTLQSGVPS RFSGSGSGTN YSLTITNLEQ 101DDAATYFCQQ GDTLPYTFGG GTKLEIKRAD AAPTVSIFPP SSEQLTSGGA 151 SVVCFLNNFYPKDINVKWKI DGSERQNGVL NSWTDQDSKD STYSMSSTLT 201 LTKDEYERHN SYTCEATHKTSTSPIVKSFN RNECNucleic acid sequence of the Ab-12 LC including signal peptide encodingsequence:

(SEQ ID NO: 200) 1 ATGATGTCCT CTGCTCAGTT CCTTGGTCTC CTGTTGCTCTGTTTTCAAGG 51 TTCCAGATGT GATCTCCAGA TGACACAGAC TACTTCCTCC CTGTCTGCCT 101CTCTGGGAGA CAGAGTCACC ATCAGTTGCA GGGCAAGTCA GGACATTAGC 151 AATTATTTAAACTGGTATCA GCAGAAACCA GATGGAACTG TTAAGCTCCT 201 GATCTTCTAC ACATCAACATTACAGTCAGG AGTCCCATCG AGGTTCAGTG 251 GCAGTGGGTC TGGAACAAAT TATTCTCTCACCATTACCAA CCTGGAGCAA 301 GATGATGCTG CCACTTACTT TTGCCAACAG GGTGATACGCTTCCGTACAC 351 GTTCGGAGGG GGGACCAAGC TGGAAATAAA ACGGGCTGAT GCTGCACCAA401 CTGTATCCAT CTTCCCACCA TCCAGTGAGC AGTTAACATC TGGAGGTGCC 451TCAGTCGTGT GCTTCTTGAA CAACTTCTAC CCCAAAGACA TCAATGTCAA 501 GTGGAAGATTGATGGCAGTG AACGACAAAA TGGCGTCCTG AACAGTTGGA 551 CTGATCAGGA CAGCAAAGACAGCACCTACA GCATGAGCAG CACCCTCACG 601 TTGACCAAGG ACGAGTATGA ACGACATAACAGCTATACCT GTGAGGCCAC 651 TCACAAGACA TCAACTTCAC CCATTGTCAA GAGCTTCAACAGGAATGAGT 701 GTTAG

Ab-12 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-12 HC:

(SEQ ID NO: 201)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-12 HC:

(SEQ ID NO: 202) 1 GAGGTCCAGT TGCAACAGTC TGGACCTGAA CTAATGAAGCCTGGGGCTTC 51 AGTGAAGATG TCCTGCAAGG CTTCTGGATA CACATTCACT GACTACAACA 101TGCACTGGAT GAAGCAGAAC CAAGGAAAGA GCCTAGAGTG GATAGGAGAG 151 ATTAATCCTAACAGTGGTGG TTCTGGTTAC AACCAGAAGT TCAAAGGCAA 201 GGCCACATTG ACTGTAGACAAGTCCTCCAG CACAGCCTAC ATGGAGCTCC 251 GCAGCCTGAC ATCTGAGGAC TCTGCAGTCTATTACTGTGC AAGATTGGGC 301 TACTATGGTA ACTACGAGGA CTGGTATTTC GATGTCTGGGGCGCAGGGAC 351 CACGGTCACC GTCTCCTCTG CCAAAACGAC ACCCCCATCT GTCTATCCAC401 TGGCCCCTGG ATCTGCTGCC CAAACTAACT CCATGGTGAC CCTGGGATGC 451CTGGTCAAGG GCTATTTCCC TGAGCCAGTG ACAGTGACCT GGAACTCTGG 501 ATCCCTGTCCAGCGGTGTGC ACACCTTCCC AGCTGTCCTG CAGTCTGACC 551 TCTACACTCT GAGCAGCTCAGTGACTGTCC CCTCCAGCAC CTGGCCCAGC 601 GAGACCGTCA CCTGCAACGT TGCCCACCCGGCCAGCAGCA CCAAGGTGGA 651 CAAGAAAATT GTGCCCAGGG ATTGTGGTTG TAAGCCTTGCATATGTACAG 701 TCCCAGAAGT ATCATCTGTC TTCATCTTCC CCCCAAAGCC CAAGGATGTG751 CTCACCATTA CTCTGACTCC TAAGGTCACG TGTGTTGTGG TAGACATCAG 801CAAGGATGAT CCCGAGGTCC AGTTCAGCTG GTTTGTAGAT GATGTGGAGG 851 TGCACACAGCTCAGACGCAA CCCCGGGAGG AGCAGTTCAA CAGCACTTTC 901 CGCTCAGTCA GTGAACTTCCCATCATGCAC CAGGACTGGC TCAATGGCAA 951 GGAGTTCAAA TGCAGGGTCA ACAGTGCAGCTTTCCCTGCC CCCATCGAGA 1001 AAACCATCTC CAAAACCAAA GGCAGACCGA AGGCTCCACAGGTGTACACC 1051 ATTCCACCTC CCAAGGAGCA GATGGCCAAG GATAAAGTCA GTCTGACCTG1101 CATGATAACA GACTTCTTCC CTGAAGACAT TACTGTGGAG TGGCAGTGGA 1151ATGGGCAGCC AGCGGAGAAC TACAAGAACA CTCAGCCCAT CATGGACACA 1201 GATGGCTCTTACTTCATCTA CAGCAAGCTC AATGTGCAGA AGAGCAACTG 1251 GGAGGCAGGA AATACTTTCACCTGCTCTGT GTTACATGAG GGCCTGCACA 1301 ACCACCATAC TGAGAAGAGC CTCTCCCACTCTCCTGGTAA ATGAAmino acid sequence of the Ab-12 HC including signal peptide:

(SEQ ID NO: 203) 1 MGWSWTFLFL LSGTSGVLSE VQLQQSGPEL MKPGASVKMSCKASGYTFTD 51 YNMHWMKQNQ GKSLEWIGEI NPNSGGSGYN QKFKGKATLT VDKSSSTAYM 101ELRSLTSEDS AVYYCARLGY YGNYEDWYFD VWGAGTTVTV SSAKTTPPSV 151 YPLAPGSAAQTNSMVTLGCL VKGYFPEPVT VTWNSGSLSS GVHTFPAVLQ 201 SDLYTLSSSV TVPSSTWPSETVTCNVAHPA SSTKVDKKIV PRDCGCKPCI 251 CTVPEVSSVF IFPPKPKDVL TITLTPKVTCVVVDISKDDP EVQFSWFVDD 301 VEVHTAQTQP REEQFNSTFR SVSELPIMHQ DWLNGKEFKCRVNSAAFPAP 351 IEKTISKTKG RPKAPQVYTI PPPKEQMAKD KVSLTCMITD FFPEDITVEW401 QWNGQPAENY KNTQPIMDTD GSYFIYSKLN VQKSNWEAGN TFTCSVLHEG 451LHNHHTEKSL SHSPGKNucleic acid sequence of the Ab-12 HC including signal peptide encodingsequence:

(SEQ ID NO: 204) 1 ATGGGATGGA GCTGGACCTT TCTCTTCCTC CTGTCAGGAACTTCGGGTGT 51 CCTCTCTGAG GTCCAGTTGC AACAGTCTGG ACCTGAACTA ATGAAGCCTG 101GGGCTTCAGT GAAGATGTCC TGCAAGGCTT CTGGATACAC ATTCACTGAC 151 TACAACATGCACTGGATGAA GCAGAACCAA GGAAAGAGCC TAGAGTGGAT 201 AGGAGAGATT AATCCTAACAGTGGTGGTTC TGGTTACAAC CAGAAGTTCA 251 AAGGCAAGGC CACATTGACT GTAGACAAGTCCTCCAGCAC AGCCTACATG 301 GAGCTCCGCA GCCTGACATC TGAGGACTCT GCAGTCTATTACTGTGCAAG 351 ATTGGGCTAC TATGGTAACT ACGAGGACTG GTATTTCGAT GTCTGGGGCG401 CAGGGACCAC GGTCACCGTC TCCTCTGCCA AAACGACACC CCCATCTGTC 451TATCCACTGG CCCCTGGATC TGCTGCCCAA ACTAACTCCA TGGTGACCCT 501 GGGATGCCTGGTCAAGGGCT ATTTCCCTGA GCCAGTGACA GTGACCTGGA 551 ACTCTGGATC CCTGTCCAGCGGTGTGCACA CCTTCCCAGC TGTCCTGCAG 601 TCTGACCTCT ACACTCTGAG CAGCTCAGTGACTGTCCCCT CCAGCACCTG 651 GCCCAGCGAG ACCGTCACCT GCAACGTTGC CCACCCGGCCAGCAGCACCA 701 AGGTGGACAA GAAAATTGTG CCCAGGGATT GTGGTTGTAA GCCTTGCATA751 TGTACAGTCC CAGAAGTATC ATCTGTCTTC ATCTTCCCCC CAAAGCCCAA 801GGATGTGCTC ACCATTACTC TGACTCCTAA GGTCACGTGT GTTGTGGTAG 851 ACATCAGCAAGGATGATCCC GAGGTCCAGT TCAGCTGGTT TGTAGATGAT 901 GTGGAGGTGC ACACAGCTCAGACGCAACCC CGGGAGGAGC AGTTCAACAG 951 CACTTTCCGC TCAGTCAGTG AACTTCCCATCATGCACCAG GACTGGCTCA 1001 ATGGCAAGGA GTTCAAATGC AGGGTCAACA GTGCAGCTTTCCCTGCCCCC 1051 ATCGAGAAAA CCATCTCCAA AACCAAAGGC AGACCGAAGG CTCCACAGGT1101 GTACACCATT CCACCTCCCA AGGAGCAGAT GGCCAAGGAT AAAGTCAGTC 1151TGACCTGCAT GATAACAGAC TTCTTCCCTG AAGACATTAC TGTGGAGTGG 1201 CAGTGGAATGGGCAGCCAGC GGAGAACTAC AAGAACACTC AGCCCATCAT 1251 GGACACAGAT GGCTCTTACTTCATCTACAG CAAGCTCAAT GTGCAGAAGA 1301 GCAACTGGGA GGCAGGAAAT ACTTTCACCTGCTCTGTGTT ACATGAGGGC 1351 CTGCACAACC ACCATACTGA GAAGAGCCTC TCCCACTCTCCTGGTAAATG 1401 A

Ab-13

The sequences of the Antibody 13 (also referred to herein as Ab-13) LCand HC are as follows:

Ab-13 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-13 LC:

(SEQ ID NO: 205)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-13 LC:

(SEQ ID NO: 206) 1 CAGATTGTTC TCACCCAGTC TCCAGCAATC ATGTCTGCATCTCCAGGGGA 51 GAAGGTCACC ATGACCTGCA GGGCCAGCTC AAGTGTAACT TCCAGTTACT 101TGAACTGGTA CCAGCAGAAG CCAGGATCTT CCCCCAAACT CTGGATTTAT 151 AGCACATCCAACCTGGCTTC AGGAGTCCCA GCTCGCTTCA GTGGCAGTGG 201 GTCTGGGACC TCTTACTCTCTCACAATCAG CAGTGTGGAG GCTGAGGATG 251 CTGCCACTTA TTACTGCCAG CAGTATGATTTTTTCCCATC GACGTTCGGT 301 GGAGGCACCA AGCTGGAAAT CAAGCGGGCT GATGCTGCACCAACTGTATC 351 CATCTTCCCA CCATCCAGTG AGCAGTTAAC ATCTGGAGGT GCCTCAGTCG401 TGTGCTTCTT GAACAACTTC TACCCCAAAG ACATCAATGT CAAGTGGAAG 451ATTGATGGCA GTGAACGACA AAATGGCGTC CTGAACAGTT GGACTGATCA 501 GGACAGCAAAGACAGCACCT ACAGCATGAG CAGCACCCTC ACGTTGACCA 551 AGGACGAGTA TGAACGACATAACAGCTATA CCTGTGAGGC CACTCACAAG 601 ACATCAACTT CACCCATCGT CAAGAGCTTCAACAGGAATG AGTGTAmino acid sequence of the Ab-13 LC including signal peptide:

(SEQ ID NO: 207) 1 MDSQVQIFSF LLISALVKMS RGQIVLTQSP AIMSASPGEKVTMTCRASSS 51 VTSSYLNWYQ QKPGSSPKLW IYSTSNLASG VPARFSGSGS GTSYSLTISS 101VEAEDAATYY CQQYDFFPST FGGGTKLEIK RADAAPTVSI FPPSSEQLTS 151 GGASVVCFLNNFYPKDINVK WKIDGSERQN GVLNSWTDQD SKDSTYSMSS 201 TLTLTKDEYE RHNSYTCEATHKTSTSPIVK SFNRNECNucleic acid sequence of the Ab-13 LC including signal peptide encodingsequence:

(SEQ ID NO: 208) 1 ATGGATTCTC AAGTGCAGAT TTTCAGCTTC CTTCTAATCAGTGCCTTAGT 51 CAAAATGTCC AGAGGACAGA TTGTTCTCAC CCAGTCTCCA GCAATCATGT 101CTGCATCTCC AGGGGAGAAG GTCACCATGA CCTGCAGGGC CAGCTCAAGT 151 GTAACTTCCAGTTACTTGAA CTGGTACCAG CAGAAGCCAG GATCTTCCCC 201 CAAACTCTGG ATTTATAGCACATCCAACCT GGCTTCAGGA GTCCCAGCTC 251 GCTTCAGTGG CAGTGGGTCT GGGACCTCTTACTCTCTCAC AATCAGCAGT 301 GTGGAGGCTG AGGATGCTGC CACTTATTAC TGCCAGCAGTATGATTTTTT 351 CCCATCGACG TTCGGTGGAG GCACCAAGCT GGAAATCAAG CGGGCTGATG401 CTGCACCAAC TGTATCCATC TTCCCACCAT CCAGTGAGCA GTTAACATCT 451GGAGGTGCCT CAGTCGTGTG CTTCTTGAAC AACTTCTACC CCAAAGACAT 501 CAATGTCAAGTGGAAGATTG ATGGCAGTGA ACGACAAAAT GGCGTCCTGA 551 ACAGTTGGAC TGATCAGGACAGCAAAGACA GCACCTACAG CATGAGCAGC 601 ACCCTCACGT TGACCAAGGA CGAGTATGAACGACATAACA GCTATACCTG 651 TGAGGCCACT CACAAGACAT CAACTTCACC CATCGTCAAGAGCTTCAACA 701 GGAATGAGTG T

Ab-13 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-13 HC:

(SEQ ID NO: 209)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-13 HC:

(SEQ ID NO: 210) 1 GAGGTCCAGC TGCAACAATC TGGACCTGAG CTGGTGAAGCCTGGGGCTTC 51 AGTGAAGATG TCCTGTAAGG CTTCTGGATA CACATTCACT GACTACTACA 101TGAACTGGGT GAAGCAGAGC CATGGAGAGA GCCTTGAGTG GATTGGAGAT 151 ATTAATCCTTACAACGATGA TACTACCTAC AACCACAAGT TCAAGGGCAA 201 GGCCACATTG ACTGTAGACAAATCCTCCAA CACAGCCTAC ATGCAGCTCA 251 ACAGCCTGAC ATCTGAGGAC TCTGCAGTCTATTACTGTGC AAGAGAGACG 301 GCCGTTATTA CTACGAATGC TATGGACTAC TGGGGTCAAGGAACCTCAGT 351 CACCGTCTCC TCAGCCAAAA CGACACCCCC ATCTGTCTAT CCACTGGCCC401 CTGGATCTGC TGCCCAAACT AACTCCATGG TGACCCTGGG ATGCCTGGTC 451AAGGGCTATT TCCCTGAGCC AGTGACAGTG ACCTGGAACT CTGGATCCCT 501 GTCCAGCGGTGTGCACACCT TCCCAGCTGT CCTGCAGTCT GACCTCTACA 551 CTCTGAGCAG CTCAGTGACTGTCCCCTCCA GCACCTGGCC CAGCGAGACC 601 GTCACCTGCA ACGTTGCCCA CCCGGCCAGCAGCACCAAGG TGGACAAGAA 651 AATTGTGCCC AGGGATTGTG GTTGTAAGCC TTGCATATGTACAGTCCCAG 701 AAGTATCATC TGTCTTCATC TTCCCCCCAA AGCCCAAGGA TGTGCTCACC751 ATTACTCTGA CTCCTAAGGT CACGTGTGTT GTGGTAGACA TCAGCAAGGA 801TGATCCCGAG GTCCAGTTCA GCTGGTTTGT AGATGATGTG GAGGTGCACA 851 CAGCTCAGACGCAACCCCGG GAGGAGCAGT TCAACAGCAC TTTCCGCTCA 901 GTCAGTGAAC TTCCCATCATGCACCAGGAC TGGCTCAATG GCAAGGAGTT 951 CAAATGCAGG GTCAACAGTG CAGCTTTCCCTGCCCCCATC GAGAAAACCA 1001 TCTCCAAAAC CAAAGGCAGA CCGAAGGCTC CACAGGTGTACACCATTCCA 1051 CCTCCCAAGG AGCAGATGGC CAAGGATAAA GTCAGTCTGA CCTGCATGAT1101 AACAGACTTC TTCCCTGAAG ACATTACTGT GGAGTGGCAG TGGAATGGGC 1151AGCCAGCGGA GAACTACAAG AACACTCAGC CCATCATGGA CACAGATGGC 1201 TCTTACTTCATCTACAGCAA GCTCAATGTG CAGAAGAGCA ACTGGGAGGC 1251 AGGAAATACT TTCACCTGCTCTGTGTTACA TGAGGGCCTG CACAACCACC 1301 ATACTGAGAA GAGCCTCTCC CACTCTCCTGGTAAAAmino acid sequence of the Ab-13 HC including signal peptide:

(SEQ ID NO: 211) 1 MGWNWIFLFL LSGTAGVYSE VQLQQSGPEL VKPGASVKMSCKASGYTFTD 51 YYMNWVKQSH GESLEWIGDI NPYNDDTTYN HKFKGKATLT VDKSSNTAYM 101QLNSLTSEDS AVYYCARETA VITTNAMDYW GQGTSVTVSS AKTTPPSVYP 151 LAPGSAAQTNSMVTLGCLVK GYFPEPVTVT WNSGSLSSGV HTFPAVLQSD 201 LYTLSSSVTV PSSTWPSETVTCNVAHPASS TKVDKKIVPR DCGCKPCICT 251 VPEVSSVFIF PPKPKDVLTI TLTPKVTCVVVDISKDDPEV QFSWFVDDVE 301 VHTAQTQPRE EQFNSTFRSV SELPIMHQDW LNGKEFKCRVNSAAFPAPIE 351 KTISKTKGRP KAPQVYTIPP PKEQMAKDKV SLTCMITDFF PEDITVEWQW401 NGQPAENYKN TQPIMDTDGS YFIYSKLNVQ KSNWEAGNTF TCSVLHEGLH 451NHHTEKSLSH SPGKNucleic acid sequence of the Ab-13 HC including signal peptide encodingsequence:

(SEQ ID NO: 212) 1 ATGGGATGGA ACTGGATCTT TCTCTTCCTC TTGTCAGGAACTGCAGGTGT 51 CTACTCTGAG GTCCAGCTGC AACAATCTGG ACCTGAGCTG GTGAAGCCTG 101GGGCTTCAGT GAAGATGTCC TGTAAGGCTT CTGGATACAC ATTCACTGAC 151 TACTACATGAACTGGGTGAA GCAGAGCCAT GGAGAGAGCC TTGAGTGGAT 201 TGGAGATATT AATCCTTACAACGATGATAC TACCTACAAC CACAAGTTCA 251 AGGGCAAGGC CACATTGACT GTAGACAAATCCTCCAACAC AGCCTACATG 301 CAGCTCAACA GCCTGACATC TGAGGACTCT GCAGTCTATTACTGTGCAAG 351 AGAGACGGCC GTTATTACTA CGAATGCTAT GGACTACTGG GGTCAAGGAA401 CCTCAGTCAC CGTCTCCTCA GCCAAAACGA CACCCCCATC TGTCTATCCA 451CTGGCCCCTG GATCTGCTGC CCAAACTAAC TCCATGGTGA CCCTGGGATG 501 CCTGGTCAAGGGCTATTTCC CTGAGCCAGT GACAGTGACC TGGAACTCTG 551 GATCCCTGTC CAGCGGTGTGCACACCTTCC CAGCTGTCCT GCAGTCTGAC 601 CTCTACACTC TGAGCAGCTC AGTGACTGTCCCCTCCAGCA CCTGGCCCAG 651 CGAGACCGTC ACCTGCAACG TTGCCCACCC GGCCAGCAGCACCAAGGTGG 701 ACAAGAAAAT TGTGCCCAGG GATTGTGGTT GTAAGCCTTG CATATGTACA751 GTCCCAGAAG TATCATCTGT CTTCATCTTC CCCCCAAAGC CCAAGGATGT 801GCTCACCATT ACTCTGACTC CTAAGGTCAC GTGTGTTGTG GTAGACATCA 851 GCAAGGATGATCCCGAGGTC CAGTTCAGCT GGTTTGTAGA TGATGTGGAG 901 GTGCACACAG CTCAGACGCAACCCCGGGAG GAGCAGTTCA ACAGCACTTT 951 CCGCTCAGTC AGTGAACTTC CCATCATGCACCAGGACTGG CTCAATGGCA 1001 AGGAGTTCAA ATGCAGGGTC AACAGTGCAG CTTTCCCTGCCCCCATCGAG 1051 AAAACCATCT CCAAAACCAA AGGCAGACCG AAGGCTCCAC AGGTGTACAC1101 CATTCCACCT CCCAAGGAGC AGATGGCCAA GGATAAAGTC AGTCTGACCT 1151GCATGATAAC AGACTTCTTC CCTGAAGACA TTACTGTGGA GTGGCAGTGG 1201 AATGGGCAGCCAGCGGAGAA CTACAAGAAC ACTCAGCCCA TCATGGACAC 1251 AGATGGCTCT TACTTCATCTACAGCAAGCT CAATGTGCAG AAGAGCAACT 1301 GGGAGGCAGG AAATACTTTC ACCTGCTCTGTGTTACATGA GGGCCTGCAC 1351 AACCACCATA CTGAGAAGAG CCTCTCCCAC TCTCCTGGTAAA

Ab-13 was humanized to generate Ab-14.

The sequences of the Antibody 14 (also referred to herein as Ab-14) LCand HC are as follows:

Ab-14 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-14 LC:

(SEQ ID NO: 213)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-14 LC:

(SEQ ID NO: 214) 1 GACATCCAGC TGACCCAGAG CCCCAGCTTC CTTTCCGCATCCGTTGGTGA 51 CCGAGTAACA ATCACATGCC GCGCCTCATC TTCAGTTACA TCTTCTTATC 101TTAATTGGTA TCAACAAAAA CCAGGAAAAG CACCTAAACT TCTTATATAC 151 TCTACATCTAATCTCGCATC AGGAGTTCCC TCTCGATTTT CAGGATCTGG 201 ATCAGGCACA GAATTTACACTTACTATATC ATCACTCCAA CCAGAAGACT 251 TCGCCACTTA TTACTGCCAA CAATACGATTTTTTTCCAAG CACATTCGGA 301 GGAGGTACAA AAGTAGAAAT CAAGCGTACG GTGGCTGCACCATCTGTCTT 351 CATCTTCCCG CCATCTGATG AGCAGTTGAA ATCTGGAACT GCCTCTGTTG401 TGTGCCTGCT GAATAACTTC TATCCCAGAG AGGCCAAAGT ACAGTGGAAG 451GTGGATAACG CCCTCCAATC GGGTAACTCC CAGGAGAGTG TCACAGAGCA 501 GGACAGCAAGGACAGCACCT ACAGCCTCAG CAGCACCCTG ACGCTGAGCA 551 AAGCAGACTA CGAGAAACACAAAGTCTACG CCTGCGAAGT CACCCATCAG 601 GGCCTGAGCT CGCCCGTCAC AAAGAGCTTCAACAGGGGAG AGTGTAmino acid sequence of the Ab-14 LC including signal peptide:

(SEQ ID NO: 215) 1 MDMRVPAQLL GLLLLWLPGA RCDIQLTQSP SFLSASVGDRVTITCRASSS 51 VTSSYLNWYQ QKPGKAPKLL IYSTSNLASG VPSRFSGSGS GTEFTLTISS 101LQPEDFATYY CQQYDFFPST FGGGTKVEIK RTVAAPSVFI FPPSDEQLKS 151 GTASVVCLLNNFYPREAKVQ WKVDNALQSG NSQESVTEQD SKDSTYSLSS 201 TLTLSKADYE KHKVYACEVTHQGLSSPVTK SFNRGECNucleic acid sequence of the Ab-14 LC including signal peptide encodingsequence:

(SEQ ID NO: 216) 1 ATGGACATGA GGGTCCCCGC TCAGCTCCTG GGGCTCCTGCTACTCTGGCT 51 CCCAGGTGCC AGATGTGACA TCCAGCTGAC CCAGAGCCCC AGCTTCCTTT 101CCGCATCCGT TGGTGACCGA GTAACAATCA CATGCCGCGC CTCATCTTCA 151 GTTACATCTTCTTATCTTAA TTGGTATCAA CAAAAACCAG GAAAAGCACC 201 TAAACTTCTT ATATACTCTACATCTAATCT CGCATCAGGA GTTCCCTCTC 251 GATTTTCAGG ATCTGGATCA GGCACAGAATTTACACTTAC TATATCATCA 301 CTCCAACCAG AAGACTTCGC CACTTATTAC TGCCAACAATACGATTTTTT 351 TCCAAGCACA TTCGGAGGAG GTACAAAAGT AGAAATCAAG CGTACGGTGG401 CTGCACCATC TGTCTTCATC TTCCCGCCAT CTGATGAGCA GTTGAAATCT 451GGAACTGCCT CTGTTGTGTG CCTGCTGAAT AACTTCTATC CCAGAGAGGC 501 CAAAGTACAGTGGAAGGTGG ATAACGCCCT CCAATCGGGT AACTCCCAGG 551 AGAGTGTCAC AGAGCAGGACAGCAAGGACA GCACCTACAG CCTCAGCAGC 601 ACCCTGACGC TGAGCAAAGC AGACTACGAGAAACACAAAG TCTACGCCTG 651 CGAAGTCACC CATCAGGGCC TGAGCTCGCC CGTCACAAAGAGCTTCAACA 701 GGGGAGAGTG T

Ab-14 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-14 HC:

(SEQ ID NO: 217)

Amino acid sequence of the mature form (signal peptide removed) of theAb-14 HC without carboxy-terminal lysine:

(SEQ ID NO: 393)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-14 HC:

(SEQ ID NO: 218) 1 GAGGTGCAGC TGGTGCAGAG CGGCGCCGAG GTCAAGAAACCTGGAGCAAG 51 CGTAAAGGTT AGTTGCAAAG CATCTGGATA CACATTTACC GACTACTACA 101TGAATTGGGT ACGACAAGCC CCTGGACAAA GACTTGAATG GATGGGAGAC 151 ATTAACCCTTATAACGACGA CACTACATAC AATCATAAAT TTAAAGGAAG 201 AGTTACAATT ACAAGAGATACATCCGCATC AACCGCCTAT ATGGAACTTT 251 CCTCATTGAG ATCTGAAGAC ACTGCTGTTTATTACTGTGC AAGAGAAACT 301 GCCGTTATTA CTACTAACGC TATGGATTAC TGGGGTCAAGGAACCACTGT 351 TACCGTCTCT AGTGCCTCCA CCAAGGGCCC ATCGGTCTTC CCCCTGGCGC401 CCTGCTCCAG GAGCACCTCC GAGAGCACAG CGGCCCTGGG CTGCCTGGTC 451AAGGACTACT TCCCCGAACC GGTGACGGTG TCGTGGAACT CAGGCGCTCT 501 GACCAGCGGCGTGCACACCT TCCCAGCTGT CCTACAGTCC TCAGGACTCT 551 ACTCCCTCAG CAGCGTGGTGACCGTGCCCT CCAGCAACTT CGGCACCCAG 601 ACCTACACCT GCAACGTAGA TCACAAGCCCAGCAACACCA AGGTGGACAA 651 GACAGTTGAG CGCAAATGTT GTGTCGAGTG CCCACCGTGCCCAGCACCAC 701 CTGTGGCAGG ACCGTCAGTC TTCCTCTTCC CCCCAAAACC CAAGGACACC751 CTCATGATCT CCCGGACCCC TGAGGTCACG TGCGTGGTGG TGGACGTGAG 801CCACGAAGAC CCCGAGGTCC AGTTCAACTG GTACGTGGAC GGCGTGGAGG 851 TGCATAATGCCAAGACAAAG CCACGGGAGG AGCAGTTCAA CAGCACGTTC 901 CGTGTGGTCA GCGTCCTCACCGTTGTGCAC CAGGACTGGC TGAACGGCAA 951 GGAGTACAAG TGCAAGGTCT CCAACAAAGGCCTCCCAGCC CCCATCGAGA 1001 AAACCATCTC CAAAACCAAA GGGCAGCCCC GAGAACCACAGGTGTACACC 1051 CTGCCCCCAT CCCGGGAGGA GATGACCAAG AACCAGGTCA GCCTGACCTG1101 CCTGGTCAAA GGCTTCTACC CCAGCGACAT CGCCGTGGAG TGGGAGAGCA 1151ATGGGCAGCC GGAGAACAAC TACAAGACCA CACCTCCCAT GCTGGACTCC 1201 GACGGCTCCTTCTTCCTCTA CAGCAAGCTC ACCGTGGACA AGAGCAGGTG 1251 GCAGCAGGGG AACGTCTTCTCATGCTCCGT GATGCATGAG GCTCTGCACA 1301 ACCACTACAC GCAGAAGAGC CTCTCCCTGTCTCCGGGTAA AAmino acid sequence of the Ab-14 HC including signal peptide:

(SEQ ID NO: 219) 1 MDWTWRILFL VAAATGAHSE VQLVQSGAEV KKPGASVKVSCKASGYTFTD 51 YYMNWVRQAP GQRLEWMGDI NPYNDDTTYN HKFKGRVTIT RDTSASTAYM 101ELSSLRSEDT AVYYCARETA VITTNAMDYW GQGTTVTVSS ASTKGPSVFP 151 LAPCSRSTSESTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS 201 GLYSLSSVVT VPSSNFGTQTYTCNVDHKPS NTKVDKTVER KCCVECPPCP 251 APPVAGPSVF LFPPKPKDTL MISRTPEVTCVVVDVSHEDP EVQFNWYVDG 301 VEVHNAKTKP REEQFNSTFR VVSVLTVVHQ DWLNGKEYKCKVSNKGLPAP 351 IEKTISKTKG QPREPQVYTL PPSREEMTKN QVSLTCLVKG FYPSDIAVEW401 ESNGQPENNY KTTPPMLDSD GSFFLYSKLT VDKSRWQQGN VFSCSVMHEA 451LHNHYTQKSL SLSPGKNucleic acid sequence of the Ab-14 HC including signal peptide encodingsequence:

(SEQ ID NO: 220) 1 ATGGACTGGA CCTGGAGGAT CCTCTTCTTG GTGGCAGCAGCCACAGGAGC 51 CCACTCCGAG GTGCAGCTGG TGCAGAGCGG CGCCGAGGTC AAGAAACCTG 101GAGCAAGCGT AAAGGTTAGT TGCAAAGCAT CTGGATACAC ATTTACCGAC 151 TACTACATGAATTGGGTACG ACAAGCCCCT GGACAAAGAC TTGAATGGAT 201 GGGAGACATT AACCCTTATAACGACGACAC TACATACAAT CATAAATTTA 251 AAGGAAGAGT TACAATTACA AGAGATACATCCGCATCAAC CGCCTATATG 301 GAACTTTCCT CATTGAGATC TGAAGACACT GCTGTTTATTACTGTGCAAG 351 AGAAACTGCC GTTATTACTA CTAACGCTAT GGATTACTGG GGTCAAGGAA401 CCACTGTTAC CGTCTCTAGT GCCTCCACCA AGGGCCCATC GGTCTTCCCC 451CTGGCGCCCT GCTCCAGGAG CACCTCCGAG AGCACAGCGG CCCTGGGCTG 501 CCTGGTCAAGGACTACTTCC CCGAACCGGT GACGGTGTCG TGGAACTCAG 551 GCGCTCTGAC CAGCGGCGTGCACACCTTCC CAGCTGTCCT ACAGTCCTCA 601 GGACTCTACT CCCTCAGCAG CGTGGTGACCGTGCCCTCCA GCAACTTCGG 651 CACCCAGACC TACACCTGCA ACGTAGATCA CAAGCCCAGCAACACCAAGG 701 TGGACAAGAC AGTTGAGCGC AAATGTTGTG TCGAGTGCCC ACCGTGCCCA751 GCACCACCTG TGGCAGGACC GTCAGTCTTC CTCTTCCCCC CAAAACCCAA 801GGACACCCTC ATGATCTCCC GGACCCCTGA GGTCACGTGC GTGGTGGTGG 851 ACGTGAGCCACGAAGACCCC GAGGTCCAGT TCAACTGGTA CGTGGACGGC 901 GTGGAGGTGC ATAATGCCAAGACAAAGCCA CGGGAGGAGC AGTTCAACAG 951 CACGTTCCGT GTGGTCAGCG TCCTCACCGTTGTGCACCAG GACTGGCTGA 1001 ACGGCAAGGA GTACAAGTGC AAGGTCTCCA ACAAAGGCCTCCCAGCCCCC 1051 ATCGAGAAAA CCATCTCCAA AACCAAAGGG CAGCCCCGAG AACCACAGGT1101 GTACACCCTG CCCCCATCCC GGGAGGAGAT GACCAAGAAC CAGGTCAGCC 1151TGACCTGCCT GGTCAAAGGC TTCTACCCCA GCGACATCGC CGTGGAGTGG 1201 GAGAGCAATGGGCAGCCGGA GAACAACTAC AAGACCACAC CTCCCATGCT 1251 GGACTCCGAC GGCTCCTTCTTCCTCTACAG CAAGCTCACC GTGGACAAGA 1301 GCAGGTGGCA GCAGGGGAAC GTCTTCTCATGCTCCGTGAT GCATGAGGCT 1351 CTGCACAACC ACTACACGCA GAAGAGCCTC TCCCTGTCTCCGGGTAAAThe CDR sequences in the variable region of the heavy chain of Ab-14are:

(SEQ ID NO: 296) CDR-H1: DYYMN (SEQ ID NO: 297) CDR-H2:DINPYNDDTTYNHKFKG (SEQ ID NO: 298) CDR-H3: ETAVITTNAMDThe light chain variable region CDR sequences of Ab-14 are:

(SEQ ID NO: 284) CDR-L1: RASSSVTSSYLN (SEQ ID NO: 285) CDR-L2: STSNLAS(SEQ ID NO: 286) CDR-L3: QQYDFFPST

Ab-14 Variable Domains:

Ab-14 light chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 380)

Ab-14 light chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 381) 1 GACATCCAGC TGACCCAGAG CCCCAGCTTC CTTTCCGCATCCGTTGGTGA 51 CCGAGTAACA ATCACATGCC GCGCCTCATC TTCAGTTACA TCTTCTTATC 101TTAATTGGTA TCAACAAAAA CCAGGAAAAG CACCTAAACT TCTTATATAC 151 TCTACATCTAATCTCGCATC AGGAGTTCCC TCTCGATTTT CAGGATCTGG 201 ATCAGGCACA GAATTTACACTTACTATATC ATCACTCCAA CCAGAAGACT 251 TCGCCACTTA TTACTGCCAA CAATACGATTTTTTTCCAAG CACATTCGGA 301 GGAGGTACAA AAGTAGAAAT CAAGAb-14 heavy chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 382)

Ab-14 heavy chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 383) 1 GAGGTGCAGC TGGTGCAGAG CGGCGCCGAG GTCAAGAAACCTGGAGCAAG 51 CGTAAAGGTT AGTTGCAAAG CATCTGGATA CACATTTACC GACTACTACA 101TGAATTGGGT ACGACAAGCC CCTGGACAAA GACTTGAATG GATGGGAGAC 151 ATTAACCCTTATAACGACGA CACTACATAC AATCATAAAT TTAAAGGAAG 201 AGTTACAATT ACAAGAGATACATCCGCATC AACCGCCTAT ATGGAACTTT 251 CCTCATTGAG ATCTGAAGAC ACTGCTGTTTATTACTGTGC AAGAGAAACT 301 GCCGTTATTA CTACTAACGC TATGGATTAC TGGGGTCAAGGAACCACTGT 351 TACCGTCTCT AGT

Ab-3 was humanized to generate Ab-15.

Ab-15

The sequences of the Antibody 15 (also referred to herein as Ab-15) LCand HC are as follows:

Ab-15 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-15 LC:

(SEQ ID NO: 221)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-15 LC:

(SEQ ID NO: 222) 1 GACATCCAGA TGACCCAGTC TCCATCCTCC CTCTCAGCATCCGTAGGCGA 51 TAGAGTTACA ATAACATGCA GCGTATCATC AACTATATCA TCAAATCATC 101TTCATTGGTT CCAACAGAAA CCCGGCAAAG CACCTAAATC ACTTATATAC 151 GGCACATCAAATCTCGCATC AGGCGTTCCT TCAAGATTTT CAGGCTCTGG 201 CTCAGGCACC GACTTTACTCTTACAATATC CTCCCTCCAA CCCGAAGACT 251 TCGCAACCTA TTACTGTCAA CAATGGTCCTCATATCCACT CACATTTGGC 301 GGCGGCACAA AAGTAGAAAT TAAACGTACG GTGGCTGCACCATCTGTCTT 351 CATCTTCCCG CCATCTGATG AGCAGTTGAA ATCTGGAACT GCCTCTGTTG401 TGTGCCTGCT GAATAACTTC TATCCCAGAG AGGCCAAAGT ACAGTGGAAG 451GTGGATAACG CCCTCCAATC GGGTAACTCC CAGGAGAGTG TCACAGAGCA 501 GGACAGCAAGGACAGCACCT ACAGCCTCAG CAGCACCCTG ACGCTGAGCA 551 AAGCAGACTA CGAGAAACACAAAGTCTACG CCTGCGAAGT CACCCATCAG 601 GGCCTGAGCT CGCCCGTCAC AAAGAGCTTCAACAGGGGAG AGTGTAmino acid sequence of the Ab-15 LC including signal peptide:

(SEQ ID NO: 223) 1 MDMRVPAQLL GLLLLWLRGA RCDIQMTQSP SSLSASVGDRVTITCSVSST 51 ISSNHLHWFQ QKPGKAPKSL IYGTSNLASG VPSRFSGSGS GTDFTLTISS 101LQPEDFATYY CQQWSSYPLT FGGGTKVEIK RTVAAPSVFI FPPSDEQLKS 151 GTASVVCLLNNFYPREAKVQ WKVDNALQSG NSQESVTEQD SKDSTYSLSS 201 TLTLSKADYE KHKVYACEVTHQGLSSPVTK SFNRGECNucleic acid sequence of the Ab-15 LC including signal peptide encodingsequence:

(SEQ ID NO: 224) 1 ATGGACATGA GGGTCCCCGC TCAGCTCCTG GGGCTCCTGCTACTCTGGCT 51 CCGAGGTGCC AGATGTGACA TCCAGATGAC CCAGTCTCCA TCCTCCCTCT 101CAGCATCCGT AGGCGATAGA GTTACAATAA CATGCAGCGT ATCATCAACT 151 ATATCATCAAATCATCTTCA TTGGTTCCAA CAGAAACCCG GCAAAGCACC 201 TAAATCACTT ATATACGGCACATCAAATCT CGCATCAGGC GTTCCTTCAA 251 GATTTTCAGG CTCTGGCTCA GGCACCGACTTTACTCTTAC AATATCCTCC 301 CTCCAACCCG AAGACTTCGC AACCTATTAC TGTCAACAATGGTCCTCATA 351 TCCACTCACA TTTGGCGGCG GCACAAAAGT AGAAATTAAA CGTACGGTGG401 CTGCACCATC TGTCTTCATC TTCCCGCCAT CTGATGAGCA GTTGAAATCT 451GGAACTGCCT CTGTTGTGTG CCTGCTGAAT AACTTCTATC CCAGAGAGGC 501 CAAAGTACAGTGGAAGGTGG ATAACGCCCT CCAATCGGGT AACTCCCAGG 551 AGAGTGTCAC AGAGCAGGACAGCAAGGACA GCACCTACAG CCTCAGCAGC 601 ACCCTGACGC TGAGCAAAGC AGACTACGAGAAACACAAAG TCTACGCCTG 651 CGAAGTCACC CATCAGGGCC TGAGCTCGCC CGTCACAAAGAGCTTCAACA 701 GGGGAGAGTG T

Ab-15 Heavy Chain

Amino acid sequence of the mature form (signal peptide removed) of Ab-15HC.

(SEQ ID NO: 225)

Amino acid sequence of the mature form (signal peptide removed) of Ab-15HC without carboxy-terminal lysine:

(SEQ ID NO: 394)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-15 HC:

(SEQ ID NO: 226) 1 GAGGTGCAGC TGGTGCAGTC TGGGGCTGAG GTGAAGAAGCCTGGGGCCTC 51 AGTGAAGGTC TCCTGCAAGG CTTCTGACTT CAACATTAAA GACTTCTATC 101TACACTGGGT GCGACAGGCC CCTGGACAAG GGCTTGAGTG GATTGGAAGG 151 ATTGATCCTGAGAATGGTGA TACTTTATAT GACCCGAAGT TCCAGGACAA 201 GGTCACCATG ACCACAGACACGTCCACCAG CACAGCCTAC ATGGAGCTGA 251 GGAGCCTGAG ATCTGACGAC ACGGCCGTGTATTACTGTGC GAGAGAGGCG 301 GATTATTTCC ACGATGGTAC CTCCTACTGG TACTTCGATGTCTGGGGCCG 351 TGGCACCCTG GTCACCGTCT CTAGTGCCTC CACCAAGGGC CCATCGGTCT401 TCCCCCTGGC GCCCTGCTCC AGGAGCACCT CCGAGAGCAC AGCGGCCCTG 451GGCTGCCTGG TCAAGGACTA CTTCCCCGAA CCGGTGACGG TGTCGTGGAA 501 CTCAGGCGCTCTGACCAGCG GCGTGCACAC CTTCCCAGCT GTCCTACAGT 551 CCTCAGGACT CTACTCCCTCAGCAGCGTGG TGACCGTGCC CTCCAGCAAC 601 TTCGGCACCC AGACCTACAC CTGCAACGTAGATCACAAGC CCAGCAACAC 651 CAAGGTGGAC AAGACAGTTG AGCGCAAATG TTGTGTCGAGTGCCCACCGT 701 GCCCAGCACC ACCTGTGGCA GGACCGTCAG TCTTCCTCTT CCCCCCAAAA751 CCCAAGGACA CCCTCATGAT CTCCCGGACC CCTGAGGTCA CGTGCGTGGT 801GGTGGACGTG AGCCACGAAG ACCCCGAGGT CCAGTTCAAC TGGTACGTGG 851 ACGGCGTGGAGGTGCATAAT GCCAAGACAA AGCCACGGGA GGAGCAGTTC 901 AACAGCACGT TCCGTGTGGTCAGCGTCCTC ACCGTTGTGC ACCAGGACTG 951 GCTGAACGGC AAGGAGTACA AGTGCAAGGTCTCCAACAAA GGCCTCCCAG 1001 CCCCCATCGA GAAAACCATC TCCAAAACCA AAGGGCAGCCCCGAGAACCA 1051 CAGGTGTACA CCCTGCCCCC ATCCCGGGAG GAGATGACCA AGAACCAGGT1101 CAGCCTGACC TGCCTGGTCA AAGGCTTCTA CCCCAGCGAC ATCGCCGTGG 1151AGTGGGAGAG CAATGGGCAG CCGGAGAACA ACTACAAGAC CACACCTCCC 1201 ATGCTGGACTCCGACGGCTC CTTCTTCCTC TACAGCAAGC TCACCGTGGA 1251 CAAGAGCAGG TGGCAGCAGGGGAACGTCTT CTCATGCTCC GTGATGCATG 1301 AGGCTCTGCA CAACCACTAC ACGCAGAAGAGCCTCTCCCT GTCTCCGGGT 1351 AAAAmino acid sequence of the Ab-15 HC including signal peptide:

(SEQ ID NO: 227) 1 MDWTWRILFL VAAATGAHSE VQLVQSGAEV KKPGASVKVSCKASDFNIKD 51 FYLHWVRQAP GQGLEWIGRI DPENGDTLYD PKFQDKVTMT TDTSTSTAYM 101ELRSLRSDDT AVYYCAREAD YFHDGTSYWY FDVWGRGTLV TVSSASTKGP 151 SVFPLAPCSRSTSESTAALG CLVKDYFPEP VTVSWNSGAL TSGVHTFPAV 201 LQSSGLYSLS SVVTVPSSNFGTQTYTCNVD HKPSNTKVDK TVERKCCVEC 251 PPCPAPPVAG PSVFLFPPKP KDTLMISRTPEVTCVVVDVS HEDPEVQFNW 301 YVDGVEVHNA KTKPREEQFN STFRVVSVLT VVHQDWLNGKEYKCKVSNKG 351 LPAPIEKTIS KTKGQPREPQ VYTLPPSREE MTKNQVSLTC LVKGFYPSDI401 AVEWESNGQP ENNYKTTPPM LDSDGSFFLY SKLTVDKSRW QQGNVFSCSV 451MHEALHNHYT QKSLSLSPGKNucleic acid sequence of the Ab-15 HC including signal peptide encodingsequence:

(SEQ ID NO: 228) 1 ATGGACTGGA CCTGGAGGAT CCTCTTCTTG GTGGCAGCAGCCACAGGAGC 51 CCACTCCGAG GTGCAGCTGG TGCAGTCTGG GGCTGAGGTG AAGAAGCCTG 101GGGCCTCAGT GAAGGTCTCC TGCAAGGCTT CTGACTTCAA CATTAAAGAC 151 TTCTATCTACACTGGGTGCG ACAGGCCCCT GGACAAGGGC TTGAGTGGAT 201 TGGAAGGATT GATCCTGAGAATGGTGATAC TTTATATGAC CCGAAGTTCC 251 AGGACAAGGT CACCATGACC ACAGACACGTCCACCAGCAC AGCCTACATG 301 GAGCTGAGGA GCCTGAGATC TGACGACACG GCCGTGTATTACTGTGCGAG 351 AGAGGCGGAT TATTTCCACG ATGGTACCTC CTACTGGTAC TTCGATGTCT401 GGGGCCGTGG CACCCTGGTC ACCGTCTCTA GTGCCTCCAC CAAGGGCCCA 451TCGGTCTTCC CCCTGGCGCC CTGCTCCAGG AGCACCTCCG AGAGCACAGC 501 GGCCCTGGGCTGCCTGGTCA AGGACTACTT CCCCGAACCG GTGACGGTGT 551 CGTGGAACTC AGGCGCTCTGACCAGCGGCG TGCACACCTT CCCAGCTGTC 601 CTACAGTCCT CAGGACTCTA CTCCCTCAGCAGCGTGGTGA CCGTGCCCTC 651 CAGCAACTTC GGCACCCAGA CCTACACCTG CAACGTAGATCACAAGCCCA 701 GCAACACCAA GGTGGACAAG ACAGTTGAGC GCAAATGTTG TGTCGAGTGC751 CCACCGTGCC CAGCACCACC TGTGGCAGGA CCGTCAGTCT TCCTCTTCCC 801CCCAAAACCC AAGGACACCC TCATGATCTC CCGGACCCCT GAGGTCACGT 851 GCGTGGTGGTGGACGTGAGC CACGAAGACC CCGAGGTCCA GTTCAACTGG 901 TACGTGGACG GCGTGGAGGTGCATAATGCC AAGACAAAGC CACGGGAGGA 951 GCAGTTCAAC AGCACGTTCC GTGTGGTCAGCGTCCTCACC GTTGTGCACC 1001 AGGACTGGCT GAACGGCAAG GAGTACAAGT GCAAGGTCTCCAACAAAGGC 1051 CTCCCAGCCC CCATCGAGAA AACCATCTCC AAAACCAAAG GGCAGCCCCG1101 AGAACCACAG GTGTACACCC TGCCCCCATC CCGGGAGGAG ATGACCAAGA 1151ACCAGGTCAG CCTGACCTGC CTGGTCAAAG GCTTCTACCC CAGCGACATC 1201 GCCGTGGAGTGGGAGAGCAA TGGGCAGCCG GAGAACAACT ACAAGACCAC 1251 ACCTCCCATG CTGGACTCCGACGGCTCCTT CTTCCTCTAC AGCAAGCTCA 1301 CCGTGGACAA GAGCAGGTGG CAGCAGGGGAACGTCTTCTC ATGCTCCGTG 1351 ATGCATGAGG CTCTGCACAA CCACTACACG CAGAAGAGCCTCTCCCTGTC 1401 TCCGGGTAAA

The CDR sequences in the variable region of the heavy chain of Ab-15are:

(SEQ ID NO: 290) CDR-H1: DFYLH (SEQ ID NO: 291) CDR-H2:RIDPENGDTLYDPKFQD (SEQ ID NO: 292) CDR-H3: EADYFHDGTSYWYFDV

The light chain variable region CDR sequences of Ab-15 are:

(SEQ ID NO: 278) CDR-L1: SVSSTISSNHLH (SEQ ID NO: 279) CDR-L2: GTSNLAS(SEQ ID NO: 280) CDR-L3: QQWSSYPLT

Ab-15 Variable Domains:

Ab-15 light chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 384)

Ab-15 light chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 385) 1 GACATCCAGA TGACCCAGTC TCCATCCTCC CTCTCAGCATCCGTAGGCGA 51 TAGAGTTACA ATAACATGCA GCGTATCATC AACTATATCA TCAAATCATC 101TTCATTGGTT CCAACAGAAA CCCGGCAAAG CACCTAAATC ACTTATATAC 151 GGCACATCAAATCTCGCATC AGGCGTTCCT TCAAGATTTT CAGGCTCTGG 201 CTCAGGCACC GACTTTACTCTTACAATATC CTCCCTCCAA CCCGAAGACT 251 TCGCAACCTA TTACTGTCAA CAATGGTCCTCATATCCACT CACATTTGGC 301 GGCGGCACAA AAGTAGAAAT TAAAAb-15 heavy chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 386)

Ab-15 heavy chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 387) 1 GAGGTGCAGC TGGTGCAGTC TGGGGCTGAG GTGAAGAAGCCTGGGGCCTC 51 AGTGAAGGTC TCCTGCAAGG CTTCTGACTT CAACATTAAA GACTTCTATC 101TACACTGGGT GCGACAGGCC CCTGGACAAG GGCTTGAGTG GATTGGAAGG 151 ATTGATCCTGAGAATGGTGA TACTTTATAT GACCCGAAGT TCCAGGACAA 201 GGTCACCATG ACCACAGACACGTCCACCAG CACAGCCTAC ATGGAGCTGA 251 GGAGCCTGAG ATCTGACGAC ACGGCCGTGTATTACTGTGC GAGAGAGGCG 301 GATTATTTCC ACGATGGTAC CTCCTACTGG TACTTCGATGTCTGGGGCCG 351 TGGCACCCTG GTCACCGTCT CTAGT

Ab-11 was humanized to generate Ab-16.

Ab-16

The sequences of the Antibody 16 (also referred to herein as Ab-16) LCand HC are as follows:

Ab-16 Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-16 LC:

(SEQ ID NO: 229)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-16 LC:

(SEQ ID NO: 230) 1 GACATCCAGT TGACCCAGTC TCCATCCTTC CTGTCTGCATCTGTAGGAGA 51 CAGAGTCACC ATCACTTGCA GGGCCAGCTC AAGTATAAGT TACATACACT 101GGTATCAGCA AAAACCAGGG AAAGCCCCTA AGCTCCTGAT CTATGCCACA 151 TCCAACCTGGCTTCTGGGGT CCCATCAAGG TTCAGCGGCA GTGGATCTGG 201 GACAGAATTC ACTCTCACAATCAGCAGCCT GCAGCCTGAA GATTTTGCAA 251 CTTATTACTG TCAGCAGTGG AGTAGTGACCCACTCACGTT CGGCGGAGGG 301 ACCAAGGTGG AGATCAAACG TACGGTGGCT GCACCATCTGTCTTCATCTT 351 CCCGCCATCT GATGAGCAGT TGAAATCTGG AACTGCCTCT GTTGTGTGCC401 TGCTGAATAA CTTCTATCCC AGAGAGGCCA AAGTACAGTG GAAGGTGGAT 451AACGCCCTCC AATCGGGTAA CTCCCAGGAG AGTGTCACAG AGCAGGACAG 501 CAAGGACAGCACCTACAGCC TCAGCAGCAC CCTGACGCTG AGCAAAGCAG 551 ACTACGAGAA ACACAAAGTCTACGCCTGCG AAGTCACCCA TCAGGGCCTG 601 AGCTCGCCCG TCACAAAGAG CTTCAACAGGGGAGAGTGTAmino acid sequence of the Ab-16 LC including signal peptide:

(SEQ ID NO: 231) 1 MDMRVPAQLL GLLLLWLPGA RCDIQLTQSP SFLSASVGDRVTITCRASSS 51 ISYIHWYQQK PGKAPKLLIY ATSNLASGVP SRFSGSGSGT EFTLTISSLQ 101PEDFATYYCQ QWSSDPLTFG GGTKVEIKRT VAAPSVFIFP PSDEQLKSGT 151 ASVVCLLNNFYPREAKVQWK VDNALQSGNS QESVTEQDSK DSTYSLSSTL 201 TLSKADYEKH KVYACEVTHQGLSSPVTKSF NRGECNucleic acid sequence of the Ab-16 LC including signal peptide encodingsequence:

(SEQ ID NO: 232) 1 ATGGACATGA GGGTCCCCGC TCAGCTCCTG GGGCTCCTGCTGCTCTGGCT 51 CCCAGGTGCC AGATGTGACA TCCAGTTGAC CCAGTCTCCA TCCTTCCTGT 101CTGCATCTGT AGGAGACAGA GTCACCATCA CTTGCAGGGC CAGCTCAAGT 151 ATAAGTTACATACACTGGTA TCAGCAAAAA CCAGGGAAAG CCCCTAAGCT 201 CCTGATCTAT GCCACATCCAACCTGGCTTC TGGGGTCCCA TCAAGGTTCA 251 GCGGCAGTGG ATCTGGGACA GAATTCACTCTCACAATCAG CAGCCTGCAG 301 CCTGAAGATT TTGCAACTTA TTACTGTCAG CAGTGGAGTAGTGACCCACT 351 CACGTTCGGC GGAGGGACCA AGGTGGAGAT CAAACGTACG GTGGCTGCAC401 CATCTGTCTT CATCTTCCCG CCATCTGATG AGCAGTTGAA ATCTGGAACT 451GCCTCTGTTG TGTGCCTGCT GAATAACTTC TATCCCAGAG AGGCCAAAGT 501 ACAGTGGAAGGTGGATAACG CCCTCCAATC GGGTAACTCC CAGGAGAGTG 551 TCACAGAGCA GGACAGCAAGGACAGCACCT ACAGCCTCAG CAGCACCCTG 601 ACGCTGAGCA AAGCAGACTA CGAGAAACACAAAGTCTACG CCTGCGAAGT 651 CACCCATCAG GGCCTGAGCT CGCCCGTCAC AAAGAGCTTCAACAGGGGAG 701 AGTGT

Ab-16 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-16 HC:

(SEQ ID NO: 233)

Amino acid sequence of the mature form (signal peptide removed) of theAb-16 HC without carboxy-terminal lysine:

(SEQ ID NO: 395)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-16 HC:

(SEQ ID NO: 234) 1 GAGGTGCAGC TGGTGCAGTC TGGGGCTGAG GTGAAGAAGCCTGGGGCCTC 51 AGTGAAGGTC TCCTGCAAGG CTTCTGGATT CGACATTAAG GACTACTATA 101TACACTGGGT GCGACAGGCC CCTGGACAAG GGCTTGAGTG GATCGGAAGG 151 GTTGATCCTGACAATGGTGA GACTGAATTT GCCCCGAAGT TCCCGGGCAA 201 GGTCACCATG ACCACAGACACGTCCATCAG CACAGCCTAC ATGGAGCTGA 251 GCAGGCTGAG ATCTGACGAC ACGGCCGTGTATTACTGTGC GAGAGAAGAC 301 TACGATGGTA CCTACACCTG GTTTCCTTAT TGGGGCCAAGGGACTCTGGT 351 CACCGTCTCT AGTGCCTCCA CCAAGGGCCC ATCGGTCTTC CCCCTGGCGC401 CCTGCTCCAG GAGCACCTCC GAGAGCACAG CGGCCCTGGG CTGCCTGGTC 451AAGGACTACT TCCCCGAACC GGTGACGGTG TCGTGGAACT CAGGCGCTCT 501 GACCAGCGGCGTGCACACCT TCCCAGCTGT CCTACAGTCC TCAGGACTCT 551 ACTCCCTCAG CAGCGTGGTGACCGTGCCCT CCAGCAACTT CGGCACCCAG 601 ACCTACACCT GCAACGTAGA TCACAAGCCCAGCAACACCA AGGTGGACAA 651 GACAGTTGAG CGCAAATGTT GTGTCGAGTG CCCACCGTGCCCAGCACCAC 701 CTGTGGCAGG ACCGTCAGTC TTCCTCTTCC CCCCAAAACC CAAGGACACC751 CTCATGATCT CCCGGACCCC TGAGGTCACG TGCGTGGTGG TGGACGTGAG 801CCACGAAGAC CCCGAGGTCC AGTTCAACTG GTACGTGGAC GGCGTGGAGG 851 TGCATAATGCCAAGACAAAG CCACGGGAGG AGCAGTTCAA CAGCACGTTC 901 CGTGTGGTCA GCGTCCTCACCGTTGTGCAC CAGGACTGGC TGAACGGCAA 951 GGAGTACAAG TGCAAGGTCT CCAACAAAGGCCTCCCAGCC CCCATCGAGA 1001 AAACCATCTC CAAAACCAAA GGGCAGCCCC GAGAACCACAGGTGTACACC 1051 CTGCCCCCAT CCCGGGAGGA GATGACCAAG AACCAGGTCA GCCTGACCTG1101 CCTGGTCAAA GGCTTCTACC CCAGCGACAT CGCCGTGGAG TGGGAGAGCA 1151ATGGGCAGCC GGAGAACAAC TACAAGACCA CACCTCCCAT GCTGGACTCC 1201 GACGGCTCCTTCTTCCTCTA CAGCAAGCTC ACCGTGGACA AGAGCAGGTG 1251 GCAGCAGGGG AACGTCTTCTCATGCTCCGT GATGCATGAG GCTCTGCACA 1301 ACCACTACAC GCAGAAGAGC CTCTCCCTGTCTCCGGGTAA AAmino acid sequence of the Ab-16 HC including signal peptide:

(SEQ ID NO: 235) 1 MDWTWRILFL VAAATGAHSE VQLVQSGAEV KKPGASVKVSCKASGFDIKD 51 YYIHWVRQAP GQGLEWIGRV DPDNGETEFA PKFPGKVTMT TDTSISTAYM 101ELSRLRSDDT AVYYCAREDY DGTYTWFPYW GQGTLVTVSS ASTKGPSVFP 151 LAPCSRSTSESTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS 201 GLYSLSSVVT VPSSNFGTQTYTCNVDHKPS NTKVDKTVER KCCVECPPCP 251 APPVAGPSVF LFPPKPKDTL MISRTPEVTCVVVDVSHEDP EVQFNWYVDG 301 VEVHNAKTKP REEQFNSTFR VVSVLTVVHQ DWLNGKEYKCKVSNKGLPAP 351 IEKTISKTKG QPREPQVYTL PPSREEMTKN QVSLTCLVKG FYPSDIAVEW401 ESNGQPENNY KTTPPMLDSD GSFFLYSKLT VDKSRWQQGN VFSCSVMHEA 451LHNHYTQKSL SLSPGKNucleic acid sequence of the Ab-16 HC including signal peptide encodingsequence:

(SEQ ID NO: 236) 1 ATGGACTGGA CCTGGAGGAT CCTCTTCTTG GTGGCAGCAGCCACAGGAGC 51 CCACTCCGAG GTGCAGCTGG TGCAGTCTGG GGCTGAGGTG AAGAAGCCTG 101GGGCCTCAGT GAAGGTCTCC TGCAAGGCTT CTGGATTCGA CATTAAGGAC 151 TACTATATACACTGGGTGCG ACAGGCCCCT GGACAAGGGC TTGAGTGGAT 201 CGGAAGGGTT GATCCTGACAATGGTGAGAC TGAATTTGCC CCGAAGTTCC 251 CGGGCAAGGT CACCATGACC ACAGACACGTCCATCAGCAC AGCCTACATG 301 GAGCTGAGCA GGCTGAGATC TGACGACACG GCCGTGTATTACTGTGCGAG 351 AGAAGACTAC GATGGTACCT ACACCTGGTT TCCTTATTGG GGCCAAGGGA401 CTCTGGTCAC CGTCTCTAGT GCCTCCACCA AGGGCCCATC GGTCTTCCCC 451CTGGCGCCCT GCTCCAGGAG CACCTCCGAG AGCACAGCGG CCCTGGGCTG 501 CCTGGTCAAGGACTACTTCC CCGAACCGGT GACGGTGTCG TGGAACTCAG 551 GCGCTCTGAC CAGCGGCGTGCACACCTTCC CAGCTGTCCT ACAGTCCTCA 601 GGACTCTACT CCCTCAGCAG CGTGGTGACCGTGCCCTCCA GCAACTTCGG 651 CACCCAGACC TACACCTGCA ACGTAGATCA CAAGCCCAGCAACACCAAGG 701 TGGACAAGAC AGTTGAGCGC AAATGTTGTG TCGAGTGCCC ACCGTGCCCA751 GCACCACCTG TGGCAGGACC GTCAGTCTTC CTCTTCCCCC CAAAACCCAA 801GGACACCCTC ATGATCTCCC GGACCCCTGA GGTCACGTGC GTGGTGGTGG 851 ACGTGAGCCACGAAGACCCC GAGGTCCAGT TCAACTGGTA CGTGGACGGC 901 GTGGAGGTGC ATAATGCCAAGACAAAGCCA CGGGAGGAGC AGTTCAACAG 951 CACGTTCCGT GTGGTCAGCG TCCTCACCGTTGTGCACCAG GACTGGCTGA 1001 ACGGCAAGGA GTACAAGTGC AAGGTCTCCA ACAAAGGCCTCCCAGCCCCC 1051 ATCGAGAAAA CCATCTCCAA AACCAAAGGG CAGCCCCGAG AACCACAGGT1101 GTACACCCTG CCCCCATCCC GGGAGGAGAT GACCAAGAAC CAGGTCAGCC 1151TGACCTGCCT GGTCAAAGGC TTCTACCCCA GCGACATCGC CGTGGAGTGG 1201 GAGAGCAATGGGCAGCCGGA GAACAACTAC AAGACCACAC CTCCCATGCT 1251 GGACTCCGAC GGCTCCTTCTTCCTCTACAG CAAGCTCACC GTGGACAAGA 1301 GCAGGTGGCA GCAGGGGAAC GTCTTCTCATGCTCCGTGAT GCATGAGGCT 1351 CTGCACAACC ACTACACGCA GAAGAGCCTC TCCCTGTCTCCGGGTAAA

The CDR sequences in the variable region of the heavy chain of Ab-16are:

(SEQ ID NO: 293) CDR-H1: DYYIH (SEQ ID NO: 294) CDR-H2:RVDPDNGETEFAPKFPG (SEQ ID NO: 295) CDR-H3: EDYDGTYTWFPY

The light chain variable region CDR sequences of Ab-16 are:

(SEQ ID NO: 281) CDR-L1: RASSSISYIH (SEQ ID NO: 282) CDR-L2: ATSNLAS(SEQ ID NO: 283) CDR-L3: QQWSSDPLT

Ab-16 Variable Domains:

Ab-16 light chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 388)

Ab-16 light chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 389) 1 GACATCCAGT TGACCCAGTC TCCATCCTTC CTGTCTGCATCTGTAGGAGA 51 CAGAGTCACC ATCACTTGCA GGGCCAGCTC AAGTATAAGT TACATACACT 101GGTATCAGCA AAAACCAGGG AAAGCCCCTA AGCTCCTGAT CTATGCCACA 151 TCCAACCTGGCTTCTGGGGT CCCATCAAGG TTCAGCGGCA GTGGATCTGG 201 GACAGAATTC ACTCTCACAATCAGCAGCCT GCAGCCTGAA GATTTTGCAA 251 CTTATTACTG TCAGCAGTGG AGTAGTGACCCACTCACGTT CGGCGGAGGG 301 ACCAAGGTGG AGATCAAAAb-16 heavy chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 390)

Ab-16 heavy chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 391) 1 GAGGTGCAGC TGGTGCAGTC TGGGGCTGAG GTGAAGAAGCCTGGGGCCTC 51 AGTGAAGGTC TCCTGCAAGG CTTCTGGATT CGACATTAAG GACTACTATA 101TACACTGGGT GCGACAGGCC CCTGGACAAG GGCTTGAGTG GATCGGAAGG 151 GTTGATCCTGACAATGGTGA GACTGAATTT GCCCCGAAGT TCCCGGGCAA 201 GGTCACCATG ACCACAGACACGTCCATCAG CACAGCCTAC ATGGAGCTGA 251 GCAGGCTGAG ATCTGACGAC ACGGCCGTGTATTACTGTGC GAGAGAAGAC 301 TACGATGGTA CCTACACCTG GTTTCCTTAT TGGGGCCAAGGGACTCTGGT 351 CACCGTCTCT AGT

Additional antibodies are referred to herein as Antibodies 17-22 (alsoreferred to herein as Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, and Ab-22). TheKappa Constant region for all VK regions of Ab-17, Ab-19, and Ab-21 isas follows:

(SEQ ID NO: 323) TDAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKS FNRNEC

The Heavy Constant Region for all VH regions of antibodies 17, 19 and 21is as follows:

(SEQ ID NO: 324) AKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK

In the following antibody amino acid sequences, the boxed-shaded aminoacids represent complement-determining regions (CDRs) and the underlinedamino acids represent signal peptide.

Ab-17

Amino acid sequence of the Ab-17 LC including signal peptide:

(SEQ ID NO: 299)

Nucleic acid sequence of the Ab-17 LC including signal peptide:

(SEQ ID NO: 300) ATGGATTTTCAGGTGCAGATTTTCAGCTTCATGCTAATCAGTGTCACAGTCATATTGTCCAGTGGAGAAATTGTGCTCACCCAGTCTCCAGCACTCATGGCTGCATCTCCAGGGGAGAAGGTCACCATCACCTGCAGTGTCAGCTCGAGTATAAGTTCCAGCAACTTACACTGGTCCCAGCAGAAGTCAGGAACCTCCCCCAAACTCTGGATTTATGGCACATCCAACCTTGCTTCTGGAGTCCCTGTTCGCTTCAGTGGCAGTGGATCTGGGACCTCTTATTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGTCAACAGTGGACTACTACGTATACGTTCGGATCGGGGACCAAGCTGGAGCTGAAACGTAmino acid sequence of the Ab-17 HC including signal peptide:

(SEQ ID NO: 301)

Nucleic acid sequence of the Ab-17 HC including signal peptide:

(SEQ ID NO: 302) ATGGGATGGAACTGGATCATCTTCTTCCTGATGGCAGTGGTTACAGGGGTCAATTCAGAGGTGCAGTTGCGGCAGTCTGGGGCAGACCTTGTGAAGCCAGGGGCCTCAGTCAAGTTGTCCTGCACAGCTTCTGGCTTCAACATTAAAGACTACTATATACACTGGGTGAAGCAGAGGCCTGAACAGGGCCTGGAGTGGATTGGAAGGATTGATCCTGATAATGGTGAAAGTACATATGTCCCGAAGTTCCAGGGCAAGGCCACTATAACAGCAGACACATCATCCAACACAGCCTACCTACAACTCAGAAGCCTGACATCTGAGGACACTGCCATCTATTATTGTGGGAGAGAGGGGCTCGACTATGGTGACTACTATGCTGTGGACTACTGGGGTCAAGGAACCTCGGTCACAGTCTCGAGCAb-17 was humanized to generate Ab-18.

Ab-18

Amino acid sequence of the Ab-18 LC including signal peptide:

(SEQ ID NO: 303)

Nucleic acid sequence of the Ab-18 LC including signal peptide:

(SEQ ID NO: 304) ATGGATATGCGCGTGCCGGCGCAGCTGCTGGGCCTGCTGCTGCTGTGGCTGCCGGGCGCGCGCTGCGATATTCAGCTGACCCAGAGCCCGAGCTTTCTGAGCGCGAGCGTGGGCGATCGCGTGACCATTACCTGCAGCGTGAGCAGCAGCATTAGCAGCAGCAACCTGCATTGGTATCAGCAGAAACCGGGCAAAGCGCCGAAACTGCTGATTTATGGCACCAGCAACCTGGCGAGCGGCGTGCCGAGCCGCTTTAGCGGCAGCGGCAGCGGCACCGAATTTACCCTGACCATTAGCAGCCTGCAGCCGGAAGATTTTGCGACCTATTATTGCCAGCAGTGGACCACCACCTATACCTTTGGCCAGGGCACCAAACTGGAAATTAAACGTAmino acid sequence of the Ab-18 HC including signal peptide:

(SEQ ID NO: 305)

Nucleic acid sequence of the Ab-18 HC including signal peptide:

(SEQ ID NO: 306) ATGGATTGGACCTGGAGCATTCTGTTTCTGGTGGCGGCGCCGACCGGCGCGCATAGCGAAGTGCAGCTGGTGCAGAGCGGCGCGGAAGTGAAAAAACCGGGCGCGAGCGTGAAAGTGAGCTGCAAAGCGAGCGGCTTTAACATTAAAGATTATTATATTCATTGGGTGCGCCAGGCGCCGGGCCAGGGCCTGGAATGGATGGGCCGCATTGATCCGGATAACGGCGAAAGCACCTATGTGCCGAAATTTCAGGGCCGCGTGACCATGACCACCGATACCAGCACCAGCACCGCGTATATGGAACTGCGCAGCCTGCGCAGCGATGATACCGCGGTGTATTATTGCGCGCGCGAAGGCCTGGATTATGGCGATTATTATGCGGTGGATTATTGGGGCCAGGGCACCCTGGTGACCGTCTCGAGCAb-18 light chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 368)

Ab-18 light chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 369) GATATTCAGCTGACCCAGAGCCCGAGCTTTCTGAGCGCGAGCGTGGGCGATCGCGTGACCATTACCTGCAGCGTGAGCAGCAGCATTAGCAGCAGCAACCTGCATTGGTATCAGCAGAAACCGGGCAAAGCGCCGAAACTGCTGATTTATGGCACCAGCAACCTGGCGAGCGGCGTGCCGAGCCGCTTTAGCGGCAGCGGCAGCGGCACCGAATTTACCCTGACCATTAGCAGCCTGCAGCCGGAAGATTTTGCGACCTATTATTGCCAGCAGTGGACCACCACCTATACCTTTGGCCAGGGCACCAAACTGGAAATTAAACGTAb-18 heavy chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 370)

Ab-18 heavy chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 371) GAAGTGCAGCTGGTGCAGAGCGGCGCGGAAGTGAAAAAACCGGGCGCGAGCGTGAAAGTGAGCTGCAAAGCGAGCGGCTTTAACATTAAAGATTATTATATTCATTGGGTGCGCCAGGCGCCGGGCCAGGGCCTGGAATGGATGGGCCGCATTGATCCGGATAACGGCGAAAGCACCTATGTGCCGAAATTTCAGGGCCGCGTGACCATGACCACCGATACCAGCACCAGCACCGCGTATATGGAACTGCGCAGCCTGCGCAGCGATGATACCGCGGTGTATTATTGCGCGCGCGAAGGCCTGGATTATGGCGATTATTATGCGGTGGATTATTGGGGCCAGGGCACCCT GGTGACCGTCTCGAGC

Ab-19

Amino acid sequence of the Ab-19 LC including signal peptide:

(SEQ ID NO: 307)

Nucleic acid sequence of the Ab-19 LC including signal peptide:

(SEQ ID NO: 308) ATGATGTCCTCTGCTCAGTTCCTTGGTCTCCTGTTGCTCTGTTTTCAAGGTACCAGATGTGATATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCAACATCAGCTGCAGGGCAAGTCAGGACATTAGCAGTTATTTAAACTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACTCCACATCAAGATTAAACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGGACAGATTATTCTCTCACTATTAGCAACCTGGCACAAGAAGATATTGCCACTTACTTTTGCCAACAGGATATTAAGCATCCGACGTTCGGTGGAGGCACCAAGTTGGAGCTGAAACGTAmino acid sequence of the Ab-19 HC including signal peptide:

(SEQ ID NO: 309)

Nucleic acid sequence of the Ab-19 HC including signal peptide:

(SEQ ID NO: 310) ATGGAATGGATCTGGATATTTCTCTTCCTCCTGTCAGGAACTGCAGGTGTCCACTCTGAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGTAAAGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCTTCTGGGTTCACATTCACTGACTACATTATGCACTGGGTGAAGCAGAAGCCTGGGCAGGGCCTTGAGTGGATTGGATATATTAATCCTTACAATGATGATACTGAATACAATGAGAAGTTCAAAGGCAAGGCCACACTGACTTCAGACAAATCCTCCAGCACAGCCTACATGGATCTCAGCAGTCTGACCTCTGAGGGCTCTGCGGTCTATTACTGTGCAAGATCGATTTATTACTACGATGCCCCGTTTGCTTACTGGGGCCAAGGGACTC TGGTCACAGTCTCGAGCAb-19 was humanized to generate Antibody 20 (also referred to herein asAb-20) and Antibody 23 (also referred to herein as Ab-23).

Ab-20

IgG4 versionAmino acid sequence of the Ab-20 LC including signal peptide:

(SEQ ID NO: 311)

Nucleic acid sequence of the Ab-20 LC including signal peptide:

(SEQ ID NO: 312) ATGATGTCCTCTGCTCAGTTCCTTGGTCTCCTGTTGCTCTGTTTTCAAGGTACCAGATGTGATATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGTGACCGTGTCACCATCACTTGCCGCGCAAGTCAGGATATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATTCTACTTCCCGTTTGAATAGTGGGGTCCCATCACGCTTCAGTGGCAGTGGCTCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGGATATTAAACACCCTACGTTCGGTCAAGGCACCAAGGTGGAGATCAAACGTAmino acid sequence of the Ab-20 HC including signal peptide:

(SEQ ID NO: 313)

Nucleic acid sequence of the Ab-20 HC including signal peptide:

(SEQ ID NO: 349) ATGGAATGGATCTGGATATTTCTCTTCCTCCTGTCAGGAACTGCAGGTGTCCACTCTGAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCTTCTGGTTTTACCTTCACCGACTATATTATGCACTGGGTGCGTCAGGCCCCTGGTCAAGGGCTTGAGTGGATGGGCTATATCAACCCTTATAATGATGACACCGAATACAACGAGAAGTTCAAGGGCCGTGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGCGCTCTGAGGACACGGCCGTGTATTACTGTGCGCGTTCGATTTATTACTACGATGCCCCGTTTGCTTACTGGGGCCAAGGGACTC TGGTCACAGTCTCGAGC

Ab-23

IgG2 version

Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-23 LC:

(SEQ ID NO: 341)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-23 LC:

(SEQ ID NO: 342) 1 GACATCCAGA TGACCCAGTC TCCATCCTCC CTGTCTGCATCTGTAGGTGA 51 CCGTGTCACC ATCACTTGCC GCGCAAGTCA GGATATTAGC AGCTATTTAA 101ATTGGTATCA GCAGAAACCA GGGAAAGCCC CTAAGCTCCT GATCTATTCT 151 ACTTCCCGTTTGAATAGTGG GGTCCCATCA CGCTTCAGTG GCAGTGGCTC 201 TGGGACAGAT TTCACTCTCACCATCAGCAG TCTGCAACCT GAAGATTTTG 251 CAACTTACTA CTGTCAACAG GATATTAAACACCCTACGTT CGGTCAAGGC 301 ACCAAGGTGG AGATCAAACG TACGGTGGCT GCACCATCTGTCTTCATCTT 351 CCCGCCATCT GATGAGCAGT TGAAATCTGG AACTGCCTCT GTTGTGTGCC401 TGCTGAATAA CTTCTATCCC AGAGAGGCCA AAGTACAGTG GAAGGTGGAT 451AACGCCCTCC AATCGGGTAA CTCCCAGGAG AGTGTCACAG AGCAGGACAG 501 CAAGGACAGCACCTACAGCC TCAGCAGCAC CCTGACGCTG AGCAAAGCAG 551 ACTACGAGAA ACACAAAGTCTACGCCTGCG AAGTCACCCA TCAGGGCCTG 601 AGCTCGCCCG TCACAAAGAG CTTCAACAGGGGAGAGTGTAmino acid sequence of the Ab-23 LC including signal peptide:

(SEQ ID NO: 343) 1 MDMRVPAQLL GLLLLWLRGA RCDIQMTQSP SSLSASVGDRVTITCRASQD 51 ISSYLNWYQQ KPGKAPKLLI YSTSRLNSGV PSRFSGSGSG TDFTLTISSL 101QPEDFATYYC QQDIKHPTFG QGTKVEIKRT VAAPSVFIFP PSDEQLKSGT 151 ASVVCLLNNFYPREAKVQWK VDNALQSGNS QESVTEQDSK DSTYSLSSTL 201 TLSKADYEKH KVYACEVTHQGLSSPVTKSF NRGECNucleic acid sequence of the Ab-23 LC including signal peptide encodingsequence:

(SEQ ID NO: 344) 1 ATGGACATGA GGGTGCCCGC TCAGCTCCTG GGGCTCCTGCTGCTGTGGCT 51 GAGAGGTGCC AGATGTGACA TCCAGATGAC CCAGTCTCCA TCCTCCCTGT 101CTGCATCTGT AGGTGACCGT GTCACCATCA CTTGCCGCGC AAGTCAGGAT 151 ATTAGCAGCTATTTAAATTG GTATCAGCAG AAACCAGGGA AAGCCCCTAA 201 GCTCCTGATC TATTCTACTTCCCGTTTGAA TAGTGGGGTC CCATCACGCT 251 TCAGTGGCAG TGGCTCTGGG ACAGATTTCACTCTCACCAT CAGCAGTCTG 301 CAACCTGAAG ATTTTGCAAC TTACTACTGT CAACAGGATATTAAACACCC 351 TACGTTCGGT CAAGGCACCA AGGTGGAGAT CAAACGTACG GTGGCTGCAC401 CATCTGTCTT CATCTTCCCG CCATCTGATG AGCAGTTGAA ATCTGGAACT 451GCCTCTGTTG TGTGCCTGCT GAATAACTTC TATCCCAGAG AGGCCAAAGT 501 ACAGTGGAAGGTGGATAACG CCCTCCAATC GGGTAACTCC CAGGAGAGTG 551 TCACAGAGCA GGACAGCAAGGACAGCACCT ACAGCCTCAG CAGCACCCTG 601 ACGCTGAGCA AAGCAGACTA CGAGAAACACAAAGTCTACG CCTGCGAAGT 651 CACCCATCAG GGCCTGAGCT CGCCCGTCAC AAAGAGCTTCAACAGGGGAG 701 AGTGT

Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-23 HC:

(SEQ ID NO: 345)

Amino acid sequence of the mature form (signal peptide removed) of theAb-23 HC without carboxy-terminal lysine:

(SEQ ID NO: 396)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-23 HC:

(SEQ ID NO: 346) 1 GAGGTGCAGC TGGTGCAGTC TGGGGCTGAG GTGAAGAAGCCTGGGTCCTC 51 GGTGAAGGTC TCCTGCAAGG CTTCTGGTTT TACCTTCACC GACTATATTA 101TGCACTGGGT GCGTCAGGCC CCTGGTCAAG GGCTTGAGTG GATGGGCTAT 151 ATCAACCCTTATAATGATGA CACCGAATAC AACGAGAAGT TCAAGGGCCG 201 TGTCACGATT ACCGCGGACAAATCCACGAG CACAGCCTAC ATGGAGCTGA 251 GCAGCCTGCG CTCTGAGGAC ACGGCCGTGTATTACTGTGC GCGTTCGATT 301 TATTACTACG ATGCCCCGTT TGCTTACTGG GGCCAAGGGACTCTGGTCAC 351 CGTCTCTAGT GCCTCCACCA AGGGCCCATC GGTCTTCCCC CTGGCGCCCT401 GCTCCAGGAG CACCTCCGAG AGCACAGCGG CCCTGGGCTG CCTGGTCAAG 451GACTACTTCC CCGAACCGGT GACGGTGTCG TGGAACTCAG GCGCTCTGAC 501 CAGCGGCGTGCACACCTTCC CAGCTGTCCT ACAGTCCTCA GGACTCTACT 551 CCCTCAGCAG CGTGGTGACCGTGCCCTCCA GCAACTTCGG CACCCAGACC 601 TACACCTGCA ACGTAGATCA CAAGCCCAGCAACACCAAGG TGGACAAGAC 651 AGTTGAGCGC AAATGTTGTG TCGAGTGCCC ACCGTGCCCAGCACCACCTG 701 TGGCAGGACC GTCAGTCTTC CTCTTCCCCC CAAAACCCAA GGACACCCTC751 ATGATCTCCC GGACCCCTGA GGTCACGTGC GTGGTGGTGG ACGTGAGCCA 801CGAAGACCCC GAGGTCCAGT TCAACTGGTA CGTGGACGGC GTGGAGGTGC 851 ATAATGCCAAGACAAAGCCA CGGGAGGAGC AGTTCAACAG CACGTTCCGT 901 GTGGTCAGCG TCCTCACCGTTGTGCACCAG GACTGGCTGA ACGGCAAGGA 951 GTACAAGTGC AAGGTCTCCA ACAAAGGCCTCCCAGCCCCC ATCGAGAAAA 1001 CCATCTCCAA AACCAAAGGG CAGCCCCGAG AACCACAGGTGTACACCCTG 1051 CCCCCATCCC GGGAGGAGAT GACCAAGAAC CAGGTCAGCC TGACCTGCCT1101 GGTCAAAGGC TTCTACCCCA GCGACATCGC CGTGGAGTGG GAGAGCAATG 1151GGCAGCCGGA GAACAACTAC AAGACCACAC CTCCCATGCT GGACTCCGAC 1201 GGCTCCTTCTTCCTCTACAG CAAGCTCACC GTGGACAAGA GCAGGTGGCA 1251 GCAGGGGAAC GTCTTCTCATGCTCCGTGAT GCATGAGGCT CTGCACAACC 1301 ACTACACGCA GAAGAGCCTC TCCCTGTCTCCGGGTAAAAmino acid sequence of the Ab-23 HC including signal peptide:

(SEQ ID NO: 347) 1 MDWTWRILFL VAAATGAHSE VQLVQSGAEV KKPGSSVKVSCKASGFTFTD 51 YIMHWVRQAP GQGLEWMGYI NPYNDDTEYN EKFKGRVTIT ADKSTSTAYM 101ELSSLRSEDT AVYYCARSIY YYDAPFAYWG QGTLVTVSSA STKGPSVFPL 151 APCSRSTSESTAALGCLVKD YFPEPVTVSW NSGALTSGVH TFPAVLQSSG 201 LYSLSSVVTV PSSNFGTQTYTCNVDHKPSN TKVDKTVERK CCVECPPCPA 251 PPVAGPSVFL FPPKPKDTLM ISRTPEVTCVVVDVSHEDPE VQFNWYVDGV 301 EVHNAKTKPR EEQFNSTFRV VSVLTVVHQD WLNGKEYKCKVSNKGLPAPI 351 EKTISKTKGQ PREPQVYTLP PSREEMTKNQ VSLTCLVKGF YPSDIAVEWE401 SNGQPENNYK TTPPMLDSDG SFFLYSKLTV DKSRWQQGNV FSCSVMHEAL 451HNHYTQKSLS LSPGKNucleic acid sequence of the Ab-23 HC including signal peptide encodingsequence:

(SEQ ID NO: 348) 1 ATGGACTGGA CCTGGAGGAT CCTCTTCTTG GTGGCAGCAGCCACAGGAGC 51 CCACTCCGAG GTGCAGCTGG TGCAGTCTGG GGCTGAGGTG AAGAAGCCTG 101GGTCCTCGGT GAAGGTCTCC TGCAAGGCTT CTGGTTTTAC CTTCACCGAC 151 TATATTATGCACTGGGTGCG TCAGGCCCCT GGTCAAGGGC TTGAGTGGAT 201 GGGCTATATC AACCCTTATAATGATGACAC CGAATACAAC GAGAAGTTCA 251 AGGGCCGTGT CACGATTACC GCGGACAAATCCACGAGCAC AGCCTACATG 301 GAGCTGAGCA GCCTGCGCTC TGAGGACACG GCCGTGTATTACTGTGCGCG 351 TTCGATTTAT TACTACGATG CCCCGTTTGC TTACTGGGGC CAAGGGACTC401 TGGTCACCGT CTCTAGTGCC TCCACCAAGG GCCCATCGGT CTTCCCCCTG 451GCGCCCTGCT CCAGGAGCAC CTCCGAGAGC ACAGCGGCCC TGGGCTGCCT 501 GGTCAAGGACTACTTCCCCG AACCGGTGAC GGTGTCGTGG AACTCAGGCG 551 CTCTGACCAG CGGCGTGCACACCTTCCCAG CTGTCCTACA GTCCTCAGGA 601 CTCTACTCCC TCAGCAGCGT GGTGACCGTGCCCTCCAGCA ACTTCGGCAC 651 CCAGACCTAC ACCTGCAACG TAGATCACAA GCCCAGCAACACCAAGGTGG 701 ACAAGACAGT TGAGCGCAAA TGTTGTGTCG AGTGCCCACC GTGCCCAGCA751 CCACCTGTGG CAGGACCGTC AGTCTTCCTC TTCCCCCCAA AACCCAAGGA 801CACCCTCATG ATCTCCCGGA CCCCTGAGGT CACGTGCGTG GTGGTGGACG 851 TGAGCCACGAAGACCCCGAG GTCCAGTTCA ACTGGTACGT GGACGGCGTG 901 GAGGTGCATA ATGCCAAGACAAAGCCACGG GAGGAGCAGT TCAACAGCAC 951 GTTCCGTGTG GTCAGCGTCC TCACCGTTGTGCACCAGGAC TGGCTGAACG 1001 GCAAGGAGTA CAAGTGCAAG GTCTCCAACA AAGGCCTCCCAGCCCCCATC 1051 GAGAAAACCA TCTCCAAAAC CAAAGGGCAG CCCCGAGAAC CACAGGTGTA1101 CACCCTGCCC CCATCCCGGG AGGAGATGAC CAAGAACCAG GTCAGCCTGA 1151CCTGCCTGGT CAAAGGCTTC TACCCCAGCG ACATCGCCGT GGAGTGGGAG 1201 AGCAATGGGCAGCCGGAGAA CAACTACAAG ACCACACCTC CCATGCTGGA 1251 CTCCGACGGC TCCTTCTTCCTCTACAGCAA GCTCACCGTG GACAAGAGCA 1301 GGTGGCAGCA GGGGAACGTC TTCTCATGCTCCGTGATGCA TGAGGCTCTG 1351 CACAACCACT ACACGCAGAA GAGCCTCTCC CTGTCTCCGGGTAAA

The CDR (complementarity determining region) sequences in the variableregion of the heavy chain of Ab-23 are as follows:

(SEQ ID NO: 269) CDR-H1: DYIMH (SEQ ID NO: 270) CDR-H2:YINPYNDDTEYNEKFKG (SEQ ID NO: 271) CDR-H3: SIYYYDAPFAY

The light chain variable region CDR sequences of Ab-23 are:

(SEQ ID NO: 239) CDR-L1: RASQDISSYLN (SEQ ID NO: 240) CDR-L2: STSRLNS(SEQ ID NO: 241) CDR-L3: QQDIKHPTAb-23 Variable domains:Ab-23 light chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 364) DIQMTQSPSS LSASVGDRVT ITCRASQDIS SYLNWYQQKP GKAPKLLIYSTSRLNSGVPS RFSGSGSGTD FTLTISSLQP EDFATYYCQQ DIKHPTFGQG TKVEIKAb-23 light chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 365) GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGTGACCGTGTC ACCATCACTTGCC GCGCAAGTCA GGATATTAGC AGCTATTTAAATTGGTATCAGCAGAAACCA GGGAAAGCCC CTAAGCTCCT GATCTATTCTACTTCCCGTT TGAATAGTGGGGTCCCATCA CGCTTCAGTG GCAGTGGCTCTGGGACAGAT TTCACTCTCA CCATCAGCAGTCTGCAACCT GAAGATTTTGCAACTTACTA CTGTCAACAG GATATTAAAC ACCCTACGTTCGGTCAAGGCACCAAGGTGG AGATCAAAAb-23 heavy chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 366) EVQLVQSGAE VKKPGSSVKV SCKASGFTFT DYIMHWVRQAPGQGLEWMGYINPYNDDTEY NEKFKGRVTI TADKSTSTAY MELSSLRSEDTAVYYCARSIYYYDAPFAYW GQGTLVTVSSAb-23 heavy chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 367) GAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAA GGTC TCCTGCAAGG CTTCTGGTTT TACCTTCACC GACTATATTATGCACTGGGTGCGTCAGGCC CCTGGTCAAG GGCTTGAGTG GATGGGCTATATCAACCCTT ATAATGATGACACCGAATAC AACGAGAAGT TCAAGGGCCGTGTCACGATT ACCGCGGACA AATCCACGAGCACAGCCTAC ATGGAGCTGAGCAGCCTGCG CTCTGAGGAC ACGGCCGTGT ATTACTGTGCGCGTTCGATTTATTACTACG ATGCCCCGTT TGCTTACTGGGGCCAAGGGACTCTGGTCACCGTCTCTAGT

Ab-21

Amino acid sequence of the Ab-21 LC including signal peptide:

(SEQ ID NO: 315)

Nucleic acid sequence of the Ab-21 LC including signal peptide:

(SEQ ID NO: 316) ATGAAGTCACAGACCCAGGTCTTTGTATACATGTTGCTGTGGTTGTCTGGTGTTGAAGGAGACATTGTGATGACCCAGTCTCACAAATTCATGTCCACGTCAGTAGGAGACAGGGTCACCATCACCTGCAAGGCCAGTCAGGATGTCTTTACTGCTGTAGCCTGGTATCAACAGAAACCAGGACAATCTCCTAAACTACTGATTTACTGGGCATCCACCCGGCACACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATTAGCAATGTGCAGTCTGAAGACTTGGCAGATTATTTCTGTCAACAATATAGCAGCTATCCTCTCACGTTCGGTGCTGGGACCAAGTTGGAGCTGAAACGTAmino acid sequence of the Ab-21 HC including signal peptide:

(SEQ ID NO: 317)

Nucleic acid sequence of the Ab-21 HC including signal peptide:

(SEQ ID NO: 318) ATGGGATGGAACTGGATCATCTTCTTCCTGATGGCAGTGGTTACAGGGGTCAATTCAGAGGTTCAGCTGCAGCAGTCTGGGGCTGAGCTTGTGAGGCCAGGGGCCTTAGTCAAGTTGTCCTGCAAAGCTTCTGGCTTCAATATTAAAGACTACTATATGCACTGGGTGAAGCAGAGGCCTGAACAGGGCCTGGAGTGGATTGGAAGGATTGATCCTGAGAATGGTGATATTATATATGACCCGAAGTTCCAGGGCAAGGCCAGTATAACAACAGACACATCCTCCAACACAGCCTACCTGCAGCTCAGCAGCCTGACGTCTGAGGACACTGCCGTCTATTACTGTGCTTACGATGCTGGTGACCCCGCCTGGTTTACTTACTGGGGCCAAGG GACTCTGGTCACCGTCTCGAGCAb-21 was humanized to yield Ab-22.

Ab-22

Amino acid sequence of the Ab-22 LC including signal peptide:

(SEQ ID NO: 319)

Nucleic acid sequence of the Ab-22 LC including signal peptide:

(SEQ ID NO: 320) ATGGATATGCGCGTGCCGGCGCAGCTGCTGGGCCTGCTGCTGCTGTGGCTGCGCGGCGCGCGCTGCGATATCCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGCGATCGCGTGACCATTACCTGCAAAGCGAGCCAGGATGTGTTTACCGCGGTGGCGTGGTATCAGCAGAAACCGGGCAAAGCGCCGAAACTGCTGATTTATTGGGCGAGCACCCGCCATACCGGCGTGCCGAGTCGCTTTAGCGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCAGCCTGCAGCCGGAAGATTTTGCGACCTATTATTGCCAGCAGTATAGCAGCTATCCGCTGACCTTTGGCGGCGGCACCAAAGTGGAAATTAAACGTAmino acid sequence of the Ab-22 HC including signal peptide:

(SEQ ID NO: 321)

Nucleic acid sequence of the Ab-22 HC including signal peptide:

(SEQ ID NO: 322) ATGGATTGGACCTGGAGCATTCTGTTTCTGGTGGCGGCGCCGACCGGCGCGCATAGCGAAGTGCAGCTGGTGCAGAGCGGCGCGGAAGTGAAAAAACCGGGCGCGAGCGTGAAAGTGAGCTGCAAAGCGAGCGGCTTTAACATTAAAGATTATTATATGCATTGGGTGCGCCAGGCGCCGGGCCAGGGCCTGGAATGGATCGGCCGCATTGATCCGGAAAACGGCGATATTATTTATGATCCGAAATTTCAGGGCCGCGTGACCATGACCACCGATACCAGCACCAGCACCGCGTATATGGAACTGCGCAGCCTGCGCAGCGATGATACCGCGGTGTATTATTGCGCGTATGATGCGGGCGATCCGGCGTGGTTTACCTATTGGGGCCAGGG CACCCTGGTGACCGTCTCGAGCAb-22 light chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 336) DIQMTQSPSS LSASVGDRVT ITCKASQDVF TAVAWYQQKP GKAPKLLIYWASTRHTGVPS RFSGSGSGTD FTLTISSLQP EDFATYYCQQ YSSYPLTFGG GTKVEIKRAb-22 light chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 337) GATATCCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGCGATCGCGTGACCATTACCTGCAAAGCGAGCCAGGATGTGTTTACCGCGGTGGCGTGGTATCAGCAGAAACCGGGCAAAGCGCCGAAACTGCTGATTTATTGGGCGAGCACCCGCCATACCGGCGTGCCGAGTCGCTTTAGCGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCAGCCTGCAGCCGGAAGATTTTGCGACCTATTATTGCCAGCAGTATAGCAGCTATCCGCTGACCTTTGGCGGCGGCACCAAAGTGGAAATTAAACGTAb-22 heavy chain variable domain amino acid sequence (without signalsequence):

(SEQ ID NO: 338) EVQLVQSGAE VKKPGASVKV SCKASGFNIK DYYMHWVRQAPGQGLEWIGRIDPENGDIIY DPKFQGRVTM TTDTSTSTAY MELRSLRSDDTAVYYCAYDAGDPAWFTYWG QGTLVTVSSAb-22 heavy chain variable domain DNA sequence (without signalsequence):

(SEQ ID NO: 339) GAAGTGCAGCTGGTGCAGAGCGGCGCGGAAGTGAAAAAACCGGGCGCGAGCGTGAAAGTGAGCTGCAAAGCGAGCGGCTTTAACATTAAAGATTATTATATGCATTGGGTGCGCCAGGCGCCGGGCCAGGGCCTGGAATGGATCGGCCGCATTGATCCGGAAAACGGCGATATTATTTATGATCCGAAATTTCAGGGCCGCGTGACCATGACCACCGATACCAGCACCAGCACCGCGTATATGGAACTGCGCAGCCTGCGCAGCGATGATACCGCGGTGTATTATTGCGCGTATGATGCGGGCGATCCGGCGTGGTTTACCTATTGGGGCCAGGGCACCCTGGTGACCGT CTCGAGC.For Ab-18, Ab-20, and Ab-22, the light chain human kappa constant regionis as follows:

(SEQ ID NO: 325) TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKS FNRGEC*and the heavy chain human gamma-4 constant region is as follows:

(SEQ ID NO: 326) ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK*The hinge region contains the Ser-241-Pro mutation to improve hingestability (Angal S et al, (1993), Mol Immunol, 30(1), 105-108).

Ab-24

The sequences of Antibody 24 (also referred to herein as Ab-24) LC andHC are as follows:

Light Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-24 LC:

(SEQ ID NO: 350)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-24 LC:

(SEQ ID NO: 354) 1 GACATTGTGT TGACCCAGTC TCCAGCTTCT TTGGCTGTGTCTCTAGGGCA 51 GAGGGCCACC ATCGCCTGCA AGGCCAGCCA AAGTGTTGAT TATGATGGTA 101CTAGTTATAT GAATTGGTAC CAACAGAAAC CAGGACAGCC ACCCAAACTC 151 CTCATCTATGCTGCATCCAA TCTAGAATCT GAGATCCCAG CCAGGTTTAG 201 TGGCACTGGG TCTGGGACAGACTTCACCCT CAACATCCAT CCTGTGGAGG 251 AGGAGGATAT CACAACCTAT TACTGTCAGCAAAGTAATGA GGATCCGTTC 301 ACGTTCGGAG GGGGGACCAA GTTGGAAATA AAACGGGCTGATGCTGCACC 351 AACTGTATCC ATCTTCCCAC CATCCAGTGA GCAGTTAACA TCTGGAGGTG401 CCTCAGTCGT GTGCTTCTTG AACAACTTCT ACCCCAAAGA CATCAATGTC 451AAGTGGAAGA TTGATGGCAG TGAACGACAA AATGGCGTCC TGAACAGTTG 501 GACTGATCAGGACAGCAAAG ACAGCACCTA CAGCATGAGC AGCACCCTCA 551 CGTTGACCAA GGACGAGTATGAACGACATA ACAGCTATAC CTGTGAGGCC 601 ACTCACAAGA CATCAACTTC ACCCATTGTCAAGAGCTTCA ACAGGAATGA 651 GTGTTAGAmino acid sequence of the Ab-24 LC including signal peptide:

(SEQ ID NO: 355) 1 METDTILLWV LLLWVPGSTG DIVLTQSPAS LAVSLGQRATIACKASQSVD 51 YDGTSYMNWY QQKPGQPPKL LIYAASNLES EIPARFSGTG SGTDFTLNIH 101PVEEEDITTY YCQQSNEDPF TFGGGTKLEI KRADAAPTVS IFPPSSEQLT 151 SGGASVVCFLNNFYPKDINV KWKIDGSERQ NGVLNSWTDQ DSKDSTYSMS 201 STLTLTKDEY ERHNSYTCEATHKTSTSPIV KSFNRNECNucleic acid sequence of the Ab-24 LC including signal peptide encodingsequence:

(SEQ ID NO: 356) 1 ATGGAGACAG ACACAATCCT GCTATGGGTG CTGCTGCTCTGGGTTCCAGG 51 CTCCACTGGT GACATTGTGT TGACCCAGTC TCCAGCTTCT TTGGCTGTGT 101CTCTAGGGCA GAGGGCCACC ATCGCCTGCA AGGCCAGCCA AAGTGTTGAT 151 TATGATGGTACTAGTTATAT GAATTGGTAC CAACAGAAAC CAGGACAGCC 201 ACCCAAACTC CTCATCTATGCTGCATCCAA TCTAGAATCT GAGATCCCAG 251 CCAGGTTTAG TGGCACTGGG TCTGGGACAGACTTCACCCT CAACATCCAT 301 CCTGTGGAGG AGGAGGATAT CACAACCTAT TACTGTCAGCAAAGTAATGA 351 GGATCCGTTC ACGTTCGGAG GGGGGACCAA GTTGGAAATA AAACGGGCTG401 ATGCTGCACC AACTGTATCC ATCTTCCCAC CATCCAGTGA GCAGTTAACA 451TCTGGAGGTG CCTCAGTCGT GTGCTTCTTG AACAACTTCT ACCCCAAAGA 501 CATCAATGTCAAGTGGAAGA TTGATGGCAG TGAACGACAA AATGGCGTCC 551 TGAACAGTTG GACTGATCAGGACAGCAAAG ACAGCACCTA CAGCATGAGC 601 AGCACCCTCA CGTTGACCAA GGACGAGTATGAACGACATA ACAGCTATAC 651 CTGTGAGGCC ACTCACAAGA CATCAACTTC ACCCATTGTCAAGAGCTTCA 701 ACAGGAATGA GTGTTAG

Ab-24 Heavy Chain:

Amino acid sequence of the mature form (signal peptide removed) of theAb-24 HC:

(SEQ ID NO: 357)

Nucleic acid sequence encoding the mature form (signal peptide removed)of the Ab-24 HC:

(SEQ ID NO: 361) 1 CAGGTCCAAC TACAGCAGCC TGGGACTGAG CTGGTGAGGCCTGGAACTTC 51 AGTGAAGTTG TCCTGTAAGG CTTCTGGCTA CATCTTCACC ACCTACTGGA 101TGAACTGGGT GAAACAGAGG CCTGGACAAG GCCTTGAGTG GATTGGCATG 151 ATTCATCCTTCCGCAAGTGA AATTAGGTTG GATCAGAAAT TCAAGGACAA 201 GGCCACATTG ACTCTTGACAAATCCTCCAG CACAGCCTAT ATGCACCTCA 251 GCGGCCCGAC ATCTGTGGAT TCTGCGGTCTATTACTGTGC AAGATCAGGG 301 GAATGGGGGT CTATGGACTA CTGGGGTCAA GGAACCTCAGTCACCGTCTC 351 CTCAGCCAAA ACGACACCCC CATCTGTCTA TCCACTGGCC CCTGGATCTG401 CTGCCCAAAC TAACTCCATG GTGACCCTGG GATGCCTGGT CAAGGGCTAT 451TTCCCTGAGC CAGTGACAGT GACCTGGAAC TCTGGATCCC TGTCCAGCGG 501 TGTGCACACCTTCCCAGCTG TCCTGCAGTC TGACCTCTAC ACTCTGAGCA 551 GCTCAGTGAC TGTCCCCTCCAGCACCTGGC CCAGCGAGAC CGTCACCTGC 601 AACGTTGCCC ACCCGGCCAG CAGCACCAAGGTGGACAAGA AAATTGTGCC 651 CAGGGATTGT GGTTGTAAGC CTTGCATATG TACAGTCCCAGAAGTATCAT 701 CTGTCTTCAT CTTCCCCCCA AAGCCCAAGG ATGTGCTCAC CATTACTCTG751 ACTCCTAAGG TCACGTGTGT TGTGGTAGAC ATCAGCAAGG ATGATCCCGA 801GGTCCAGTTC AGCTGGTTTG TAGATGATGT GGAGGTGCAC ACAGCTCAGA 851 CGCAACCCCGGGAGGAGCAG TTCAACAGCA CTTTCCGCTC AGTCAGTGAA 901 CTTCCCATCA TGCACCAGGACTGGCTCAAT GGCAAGGAGT TCAAATGCAG 951 GGTCAACAGT GCAGCTTTCC CTGCCCCCATCGAGAAAACC ATCTCCAAAA 1001 CCAAAGGCAG ACCGAAGGCT CCACAGGTGT ACACCATTCCACCTCCCAAG 1051 GAGCAGATGG CCAAGGATAA AGTCAGTCTG ACCTGCATGA TAACAGACTT1101 CTTCCCTGAA GACATTACTG TGGAGTGGCA GTGGAATGGG CAGCCAGCGG 1151AGAACTACAA GAACACTCAG CCCATCATGG ACACAGATGG CTCTTACTTC 1201 ATCTACAGCAAGCTCAATGT GCAGAAGAGC AACTGGGAGG CAGGAAATAC 1251 TTTCACCTGC TCTGTGTTACATGAGGGCCT GCACAACCAC CATACTGAGA 1301 AGAGCCTCTC CCACTCTCCT GGTAAATGAAmino acid sequence of the Ab-24 HC including signal peptide:

(SEQ ID NO: 362) 1 MGWSSIILFL VATATGVHSQ VQLQQPGTEL VRPGTSVKLSCKASGYIFTT 51 YWMNWVKQRP GQGLEWIGMI HPSASEIRLD QKFKDKATLT LDKSSSTAYM 101HLSGPTSVDS AVYYCARSGE WGSMDYWGQG TSVTVSSAKT TPPSVYPLAP 151 GSAAQTNSMVTLGCLVKGYF PEPVTVTWNS GSLSSGVHTF PAVLQSDLYT 201 LSSSVTVPSS TWPSETVTCNVAHPASSTKV DKKIVPRDCG CKPCICTVPE 251 VSSVFIFPPK PKDVLTITLT PKVTCVVVDISKDDPEVQFS WFVDDVEVHT 301 AQTQPREEQF NSTFRSVSEL PIMHQDWLNG KEFKCRVNSAAFPAPIEKTI 351 SKTKGRPKAP QVYTIPPPKE QMAKDKVSLT CMITDFFPED ITVEWQWNGQ401 PAENYKNTQP IMDTDGSYFI YSKLNVQKSN WEAGNTFTCS VLHEGLHNHH 451TEKSLSHSPG KNucleic acid sequence of the Ab-24 HC including signal peptide encodingsequence:

(SEQ ID NO: 363) 1 ATGGGATGGA GCTCTATCAT CCTCTTCTTG GTAGCAACAGCTACAGGTGT 51 CCACTCCCAG GTCCAACTAC AGCAGCCTGG GACTGAGCTG GTGAGGCCTG 101GAACTTCAGT GAAGTTGTCC TGTAAGGCTT CTGGCTACAT CTTCACCACC 151 TACTGGATGAACTGGGTGAA ACAGAGGCCT GGACAAGGCC TTGAGTGGAT 201 TGGCATGATT CATCCTTCCGCAAGTGAAAT TAGGTTGGAT CAGAAATTCA 251 AGGACAAGGC CACATTGACT CTTGACAAATCCTCCAGCAC AGCCTATATG 301 CACCTCAGCG GCCCGACATC TGTGGATTCT GCGGTCTATTACTGTGCAAG 351 ATCAGGGGAA TGGGGGTCTA TGGACTACTG GGGTCAAGGA ACCTCAGTCA401 CCGTCTCCTC AGCCAAAACG ACACCCCCAT CTGTCTATCC ACTGGCCCCT 451GGATCTGCTG CCCAAACTAA CTCCATGGTG ACCCTGGGAT GCCTGGTCAA 501 GGGCTATTTCCCTGAGCCAG TGACAGTGAC CTGGAACTCT GGATCCCTGT 551 CCAGCGGTGT GCACACCTTCCCAGCTGTCC TGCAGTCTGA CCTCTACACT 601 CTGAGCAGCT CAGTGACTGT CCCCTCCAGCACCTGGCCCA GCGAGACCGT 651 CACCTGCAAC GTTGCCCACC CGGCCAGCAG CACCAAGGTGGACAAGAAAA 701 TTGTGCCCAG GGATTGTGGT TGTAAGCCTT GCATATGTAC AGTCCCAGAA751 GTATCATCTG TCTTCATCTT CCCCCCAAAG CCCAAGGATG TGCTCACCAT 801TACTCTGACT CCTAAGGTCA CGTGTGTTGT GGTAGACATC AGCAAGGATG 851 ATCCCGAGGTCCAGTTCAGC TGGTTTGTAG ATGATGTGGA GGTGCACACA 901 GCTCAGACGC AACCCCGGGAGGAGCAGTTC AACAGCACTT TCCGCTCAGT 951 CAGTGAACTT CCCATCATGC ACCAGGACTGGCTCAATGGC AAGGAGTTCA 1001 AATGCAGGGT CAACAGTGCA GCTTTCCCTG CCCCCATCGAGAAAACCATC 1051 TCCAAAACCA AAGGCAGACC GAAGGCTCCA CAGGTGTACA CCATTCCACC1101 TCCCAAGGAG CAGATGGCCA AGGATAAAGT CAGTCTGACC TGCATGATAA 1151CAGACTTCTT CCCTGAAGAC ATTACTGTGG AGTGGCAGTG GAATGGGCAG 1201 CCAGCGGAGAACTACAAGAA CACTCAGCCC ATCATGGACA CAGATGGCTC 1251 TTACTTCATC TACAGCAAGCTCAATGTGCA GAAGAGCAAC TGGGAGGCAG 1301 GAAATACTTT CACCTGCTCT GTGTTACATGAGGGCCTGCA CAACCACCAT 1351 ACTGAGAAGA GCCTCTCCCA CTCTCCTGGT AAATGAThe CDR sequences in the variable region of the light chain of Ab-24 areas follows:

(SEQ ID NO: 351) CDR-L1: KASQSVDYDGTSYMN (SEQ ID NO: 352) CDR-L2:AASNLES (SEQ ID NO: 353) CDR-L3: QQSNEDPFT

The CDR sequences in the variable region of the heavy chain of Ab-24 areas follows:

(SEQ ID NO: 358) CDR-H1: TYWMN (SEQ ID NO: 359) CDR-H2:MIHPSASEIRLDQKFKD (SEQ ID NO: 360) CDR-H3: SGEWGSMDY

Table 1 below provides the SEQ ID NOs and amino acid sequences of theCDR's of Ab-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6,Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16,Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, Ab-22, Ab-23, and Ab-24. L1, L2, andL3 refer to light chain CDR's 1, 2, and 3, and H1, H2, and H3 refer toheavy chain CDR's 1, 2, and 3 according to the Kabat numbering system(Kabat et at, 1987 in Sequences of Proteins of Immunological Interest,U.S. Department of Health and Human Services, NIH, USA).

TABLE 1 SEQ ID NO DESCRIPTION AMINO ACID SEQUENCE 54 Ab-A and Ab-1CDR-L1 QSSQSVYDNNWLA 55 Ab-A and Ab-1 CDR-L2 DASDLAS 56 Ab-A and Ab-1CDR-L3 QGAYNDVIYA 51 Ab-A and Ab-1 CDR-H1 SYWMN 52 Ab-A and Ab-1 CDR-H2TIDSGGRTDYASWAKG 53 Ab-A and Ab-1 CDR-H3 NWNL 60 Ab-B CDR-L1 SASSSVSFVD61 Ab-B CDR-L2 RTSNLGF 62 Ab-B CDR-L3 QQRSTYPPT 57 Ab-B CDR-H1 TSGMGVG58 Ab-B CDR-H2 HIWWDDVKRYNPVLKS 59 Ab-B CDR-H3 EDFDYDEEYYAMDY 48 Ab-CCDR-L1 KASQSVDYDGDSYMN 49 Ab-C CDR-L2 AASNLES 50 Ab-C CDR-L3 QQSNEDPWT45 Ab-C CDR-H1 DCYMN 46 Ab-C CDR-H2 DINPFNGGTTYNQKFKG 47 Ab-C CDR-H3SHYYFDGRVPWDAMDY 42 Ab-D CDR-L1 QASQGTSINLN 43 Ab-D CDR-L2 GSSNLED 44Ab-D CDR-L3 LQHSYLPYT 39 Ab-D CDR-H1 DHYMS 40 Ab-D CDR-H2DINPYSGETTYNQKFKG 41 Ab-D CDR-H3 DDYDASPFAY 275 Ab-2 CDR-L1 RASSSVYYYMH276 Ab-2 CDR-L2 ATSNLAS 277 Ab-2 CDR-L3 QQWSSDPLT 287 Ab-2 CDR-H1 DYFIH288 Ab-2 CDR-H2 RLDPEDGESDYAPKFQD 289 Ab-2 CDR-H3 EDYDGTYTFFPY 278 Ab-3and Ab-15 CDR-L1 SVSSTISSNHLH 279 Ab-3 and Ab-15 CDR-L2 GTSNLAS 280 Ab-3and Ab-15 CDR-L3 QQWSSYPLT 290 Ab-3 and Ab-15 CDR-H1 DFYLH 291 Ab-3 andAb-15 CDR-H2 RIDPENGDTLYDPKFQD 292 Ab-3 and Ab-15 CDR-H3EADYFHDGTSYWYFDV 78 Ab-4 and Ab-5 CDR-L1 RASQDISNYLN 79 Ab-4 and Ab-5CDR-L2 YTSRLLS 80 Ab-4 and Ab-5 CDR-L3 QQGDTLPYT 245 Ab-4 and Ab-5CDR-H1 DYNMH 246 Ab-4 and Ab-5 CDR-H2 EINPNSGGAGYNQKFKG 247 Ab-4 andAb-5 CDR-H3 LGYDDIYDDWYFDV 81 Ab-6 CDR-L1 RASQDISNYLN 99 Ab-6 CDR-L2YTSRLHS 100 Ab-6 CDR-L3 QQGDTLPYT 248 Ab-6 CDR-H1 DYNMH 249 Ab-6 CDR-H2EINPNSGGSGYNQKFKG 250 Ab-6 CDR-H3 LVYDGSYEDWYFDV 101 Ab-7 CDR-L1RASQVITNYLY 102 Ab-7 CDR-L2 YTSRLHS 103 Ab-7 CDR-L3 QQGDTLPYT 251 Ab-7CDR-H1 DYNMH 252 Ab-7 CDR-H2 EINPNSGGAGYNQQFKG 253 Ab-7 CDR-H3LGYVGNYEDWYFDV 104 Ab-8 CDR-L1 RASQDISNYLN 105 Ab-8 CDR-L2 YTSRLLS 106Ab-8 CDR-L3 QQGDTLPYT 254 Ab-8 CDR-H1 DYNMH 255 Ab-8 CDR-H2EINPNSGGAGYNQKFKG 256 Ab-8 CDR-H3 LGYDDIYDDWYFDV 107 Ab-9 CDR-L1RASQDISNYLN 108 Ab-9 CDR-L2 YTSRLFS 109 Ab-9 CDR-L3 QQGDTLPYT 257 Ab-9CDR-H1 DYNMH 258 Ab-9 CDR-H2 EINPNSGGAGYNQKFKG 259 Ab-9 CDR-H3LGYDDIYDDWYFDV 110 Ab-10 CDR-L1 RASQDISNYLN 111 Ab-10 CDR-L2 YTSRLLS 112Ab-10 CDR-L3 QQGDTLPYT 260 Ab-10 CDR-H1 DYNMH 261 Ab-10 CDR-H2EINPNSGGAGYNQKFKG 262 Ab-10 CDR-H3 LGYDDIYDDWYFDV 281 Ab-11 and Ab-16CDR-L1 RASSSISYIH 282 Ab-11 and Ab-16 CDR-L2 ATSNLAS 283 Ab-11 and Ab-16CDR-L3 QQWSSDPLT 293 Ab-11 and Ab-16 CDR-H1 DYYIH 294 Ab-11 and Ab-16CDR-H2 RVDPDNGETEFAPKFPG 295 Ab-11 and Ab-16 CDR-H3 EDYDGTYTWFPY 113Ab-12 CDR-L1 RASQDISNYLN 114 Ab-12 CDR-L2 YTSTLQS 115 Ab-12 CDR-L3QQGDTLPYT 263 Ab-12 CDR-H1 DYNMH 264 Ab-12 CDR-H2 EINPNSGGSGYNQKFKG 265Ab-12 CDR-H3 LGYYGNYEDWYFDV 284 Ab-13 and Ab-14 CDR-L1 RASSSVTSSYLN 285Ab-13 and Ab-14 CDR-L2 STSNLAS 286 Ab-13 and Ab-14 CDR-L3 QQYDFFPST 296Ab-13 and Ab-14 CDR-H1 DYYMN 297 Ab-13 and Ab-14 CDR-H2DINPYNDDTTYNHKFKG 298 Ab-13 and Ab-14 CDR-H3 ETAVITTNAMD 116 Ab-17 andAb-18 CDR-L1 SVSSSISSSNLH 237 Ab-17 and Ab-18 CDR-L2 GTSNLAS 238 Ab-17and Ab-18 CDR-L3 QQWTTTYT 266 Ab-17 and Ab-18 CDR-H1 DYYIH 267 Ab-17 andAb-18 CDR-H2 RIDPDNGESTYVPKFQG 268 Ab-17 and Ab-18 CDR-H3 EGLDYGDYYAVDY239 Ab-19, Ab-20 and Ab-23 RASQDISSYLN CDR-L1 240 Ab-19, Ab-20 and Ab-23STSRLNS CDR-L2 241 Ab-19, Ab-20 and Ab-23 QQDIKHPT CDR-L3 269 Ab-19,Ab-20 and Ab-23 DYIMH CDR-H1 270 Ab-19, Ab-20 and Ab-23YINPYNDDTEYNEKFKG CDR-H2 271 Ab-19, Ab-20 and Ab-23 SIYYYDAPFAY CDR-H3242 Ab-21 and Ab-22 CDR-L1 KASQDVFTAVA 243 Ab-21 and Ab-22 CDR-L2WASTRHT 244 Ab-21 and Ab-22 CDR-L3 QQYSSYPLT 272 Ab-21 and Ab-22 CDR-H1DYYMH 273 Ab-21 and Ab-22 CDR-H2 RIDPENGDIIYDPKFQG 274 Ab-21 and Ab-22CDR-H3 DAGDPAWFTY 351 Ab-24 CDR-L1 KASQSVDYDGTSYMN 352 Ab-24 CDR-L2AASNLES 353 Ab-24 CDR-L3 QQSNEDPFT 358 Ab-24 CDR-H1 TYWMN 359 Ab-24CDR-H2 MIHPSASEIRLDQKFKD 360 Ab-24 CDR-H3 SGEWGSMDY

An oligopeptide or polypeptide is within the scope of the invention ifit has an amino acid sequence that is at least 75%, 76%, 77%, 78%, 79%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98% or 99% identical to least one of the CDR's ofTable 1 above; and/or to a CDR of a sclerostin binding agent thatcross-blocks the binding of at least one of antibodies Ab-A, Ab-B, Ab-C,Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10,Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19, Ab-20,Ab-21, Ab-22, Ab-23, and Ab-24 to sclerostin, and/or is cross-blockedfrom binding to sclerostin by at least one of antibodies Ab-A, Ab-B,Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10,Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19, Ab-20,Ab-21, Ab-22, Ab-23, and Ab-24; and/or to a CDR of a sclerostin bindingagent wherein the binding agent can block the inhibitory effect ofsclerostin in a cell based mineralization assay (i.e. a sclerostinneutralizing binding agent); and/or to a CDR of a sclerostin bindingagent that binds to a Loop 2 epitope; and/or to a CDR of a sclerostinbinding agent that binds to a T20.6 epitope; and/or to a CDR of asclerostin binding agent that binds to a “T20.6 derivative(cystine-knot+4 arms)” epitope.

Sclerostin binding agent polypeptides and antibodies are within thescope of the invention if they have amino acid sequences that are atleast 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% identical to a variable region of at least one of antibodiesAb-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8,Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18,Ab-19, Ab-20, Ab-21, Ab-22, Ab-23, and Ab-24, and cross-block thebinding of at least one of antibodies Ab-A, Ab-B, Ab-C, Ab-D, Ab-1,Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12,Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, Ab-22,Ab-23, and Ab-24 to sclerostin, and/or are cross-blocked from binding tosclerostin by at least one of antibodies Ab-A, Ab-B, Ab-C, Ab-D, Ab-1,Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12,Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, Ab-22,Ab-23, and Ab-24; and/or can block the inhibitory effect of sclerostinin a cell based mineralization assay (i.e. a sclerostin neutralizingbinding agent); and/or bind to a Loop 2 epitope; and/or bind to a T20.6epitope; and/or bind to a “T20.6 derivative (cystine-knot+4 arms)”epitope.

Polynucleotides encoding sclerostin binding agents are within the scopeof the invention if they have polynucleotide sequences that are at least85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or99% identical to a polynucleotide encoding a variable region of at leastone of antibodies Ab-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5,Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16,Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, Ab-22, Ab-23, and Ab-24, and whereinthe encoded sclerostin binding agents cross-block the binding of atleast one of antibodies Ab-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4,Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15,Ab-16, Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, Ab-22, Ab-23, and Ab-24 tosclerostin, and/or are cross-blocked from binding to sclerostin by atleast one of antibodies Ab-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4,Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15,Ab-16, Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, Ab-22, Ab-23, and Ab-24;and/or can block the inhibitory effect of sclerostin in a cell basedmineralization assay (i.e. a sclerostin neutralizing binding agent);and/or bind to a Loop 2 epitope; and/or bind to a T20.6 epitope; and/orbind to a “T20.6 derivative (cystine-knot+4 arms)” epitope.

Antibodies according to the invention may have a binding affinity forhuman sclerostin of less than or equal to 1×10⁻⁷M, less than or equal to1×10⁻⁸M, less than or equal to 1×10⁻⁹M, less than or equal to 1×10⁻¹° M,less than or equal to 1×10⁻¹¹M, or less than or equal to 1×10⁻¹²M.

The affinity of a binding agent such as an antibody or binding partner,as well as the extent to which a binding agent (such as an antibody)inhibits binding, can be determined by one of ordinary skill in the artusing conventional techniques, for example those described by Scatchardet al. (Ann. N.Y. Acad. Sci. 51:660-672 (1949)) or by surface plasmonresonance (SPR; BIAcore, Biosensor, Piscataway, N.J.). For surfaceplasmon resonance, target molecules are immobilized on a solid phase andexposed to ligands in a mobile phase running along a flow cell. Ifligand binding to the immobilized target occurs, the local refractiveindex changes, leading to a change in SPR angle, which can be monitoredin real time by detecting changes in the intensity of the reflectedlight. The rates of change of the SPR signal can be analyzed to yieldapparent rate constants for the association and dissociation phases ofthe binding reaction. The ratio of these values gives the apparentequilibrium constant (affinity) (see, e.g., Wolff et al., Cancer Res.53:2560-65 (1993)).

An antibody according to the present invention may belong to anyimmunoglobin class, for example IgG, IgE, IgM, IgD, or IgA. It may beobtained from or derived from an animal, for example, fowl (e.g.,chicken) and mammals, which includes but is not limited to a mouse, rat,hamster, rabbit, or other rodent, cow, horse, sheep, goat, camel, human,or other primate. The antibody may be an internalizing antibody.Production of antibodies is disclosed generally in U.S. PatentPublication No. 2004/0146888 A1.

Characterization Assays

In the methods described above to generate antibodies according to theinvention, including the manipulation of the specific Ab-A, Ab-B, Ab-C,Ab-D, and Antibody 1-24 (Ab-1 to Ab-24) CDRs into new frameworks and/orconstant regions, appropriate assays are available to select the desiredantibodies or binding agents (i. e. assays for determining bindingaffinity to sclerostin; cross-blocking assays; Biacore-based “humansclerostin peptide epitope competition binding assay;” MC3T3-E1 cellbased assay; in vivo assays).

Epitope Binding Assays

Mature form human sclerostin is a 190 amino acid glycoprotein with acystine-knot structure (FIGS. 8 and 9). In addition to the cystine-knotstructure, the protein is characterized as having three loops designatedas Loop 1, Loop 2 and Loop 3. Human sclerostin was subjected toproteolytic digestion to produce fragments. Briefly, using differentproteases, including trypsin, aspN, and lysC, fragments with variouscleavage sites and sizes were generated. The sequences and mass forvarious human sclerostin peptides were determined. Antibody protectionwas evaluated to determine the effect on accessibility for proteolysis,including clipped site masking and peptide shifting. Finally, aBIAcore-based “human sclerostin peptide epitope competition assay” wasperformed.

Exposure of sclerostin to trypsin cleavage resulted in a pattern ofpeptide fragments as summarized in FIG. 13. The fragments are referredto as T19.2, T20, T20.6, and T21-22. As shown schematically in FIG. 19B,the T20.6 epitope is a complex of four separate peptide sequences whichare joined by the three disulfide bonds of the cystine-knot region. Twoof the peptides are joined by two disulfide bonds. The other twopeptides are linked by one disulfide bond that, schematically, bisectsthe first two polypeptides.

The T20.6 epitope that was generated by trypsin digestion retains thecystine-knot structure of the native polypeptide and is recognized byantibodies Ab-C and Ab-D. A derivative of epitope T20.6 consists of thecystine-knot region and amino acids 58-64, 73-81, 112-117 and 138-141 insequence position with reference to SEQ ID NO:1. This derivative epitopeis shown in FIG. 21. An epitope comprising the cystine-knot region mayhave one or more amino acids that is present in the T20.6 epitope (FIG.19B) but not present in the T20.6 derivative epitope (FIG. 21).

Another epitope-containing region was identified in the Loop 2 region ofhuman sclerostin (FIG. 19A) and is recognized by antibodies Ab-A andAb-B. A Loop 2 epitope comprises amino acids 86-111 of SEQ ID NO:1(C4GPARLLPNAIGRGKWWRPSGPDFRCS, SEQ ID NO:6). Sterically, with referenceto full-length sclerostin of SEQ ID NO:1, the Loop 2-containingstructure is defined at one end by a disulfide bond between cysteine atposition 86 (C4) and cysteine at position 144 (C8), and at the other endby a disulfide bond between cysteine at position 111 (C5) and cysteineat position 57 (C1).

The peptides generated by aspN cleavage of human sclerostin are shown inFIG. 12. In the Figure, these peptides are designated as AspN14.6,AspN18.6, and AspN22.7-23.5, and are also referred to herein as N14.6,N18.6, and N22.7-23.5, respectively.

One group of antibodies exhibits a specific pattern of binding tocertain epitopes as evidenced by a Biacore-based “human sclerostinpeptide epitope competition binding assay.” Briefly, the antibody ispreincubated with the epitope to be tested, at concentrations that willsaturate the epitope-binding sites on the antibody. The antibody is thenexposed to sclerostin bound to a chip surface. After the appropriateincubation and washing procedures, a pattern of competitive binding isestablished. As shown in FIG. 18, exemplary antibody Ab-D bound tosclerostin molecules attached to the surface of the chip. Preincubationof antibody Ab-D with sclerostin decreased the binding of the antibodyto the sclerostin on the chip to close to zero. Preincubation with apeptide consisting of epitope T19.2 showed that T19.2 did not competewith sclerostin for antibody binding. However, preincubation with anyone of the epitopes designated T20, T20.6, T21-22, or N22.7-23.5abolished a large proportion of the binding of antibody to sclerostin onthe chip. In contrast, preincubation of the antibody with any one of theepitopes designated T19.2, N14.6 or N18.6 did not abolish the ability ofthe antibody to bind to sclerostin. A second exemplary antibody withthis binding profile (FIG. 17) is Ab-C.

Antibody Ab-D therefore is exemplary and representative of a group ofantibodies that bind to the epitopes T20, T20.6, T21-22, and N22.7-23.5,and have minimal detectable binding to epitopes T19.2, N14.6 and N18.6,as measured by the ability to block antibody binding to sclerostin.Antibodies having this characteristic binding pattern may or may notshare amino acid sequence in one or more regions of the antibodymolecule. Antibody similarity is determined functionally such as by theability to bind to sclerostin following preincubation with each of theepitopes described above. Antibodies that exhibit a binding patternsimilar or identical to that of antibody Ab-D are included in theinvention. By “similar to” is meant, for example, the antibody willexhibit binding to each of the polypeptides T20, T20.6, T21-22 andN22.7-23.5 whereby this binding will specifically compete out at least50% of the antibody's binding to sclerostin that would otherwise occurin the absence of preincubation with sclerostin or a sclerostin peptide.The antibody will also exhibit little or no detectable binding topolypeptides T19.2, N14.6 and N18.6, resulting in a reduction of 30% orless of the binding that would occur in the absence of preincubationwith sclerostin or a sclerostin peptide.

For example, without being bound by a particular mechanism, the antibodybinding pattern of FIG. 18 suggests that the epitope space to whichantibody Ab-D and other antibodies having the epitope binding pattern ofAb-D bind consists of a polypeptide comprising the cystine-knot regionof sclerostin.

Thus, as disclosed herein and with reference to FIG. 19B, an exemplaryT20.6 epitope comprises four peptide chains attached via three separatedisulfide bonds. Peptide chain SAKPVTELVC3SGQC4GPAR (SEQ ID NO:3) isattached to peptide chain LVASC7KC8KRLTR (SEQ ID NO:5) by disulfidebonds from C3 to C7, and from C4 to C8. Peptide chain DVSEYSC1RELHFTR(SEQ ID NO:2) is attached to peptide chain WWRPSGPDFRC5IPDRYR (SEQ IDNO:4) by a disulfide bond from C1 to C5. The polypeptides of SEQ IDNOs:3 and 5 remain associated with the polypeptides of SEQ ID NOs:2 and4 through a steric construct whereby the C1-C5 bond crosses the plane ofthe C4-C8 and C3-C7 bonds and is located between them, as illustrated inFIG. 19B.

As disclosed herein and with reference to FIG. 21, an exemplaryderivative epitope of T20.6 comprises four peptide chains attached viathree separate disulfide bonds. Peptide chain SAKPVTELVC3SGQC4 (SEQ IDNO:70) is attached to peptide chain LVASC7KC8 (SEQ ID NO:71) bydisulfide bonds from C3 to C7, and from C4 to C8. Peptide chainC1RELHFTR (SEQ ID NO:72) is attached to peptide chain C5IPDRYR (SEQ IDNO:73) by a disulfide bond from C1 to C5. The polypeptides of SEQ IDNOs:70 and 71 remain associated with the polypeptides of SEQ ID NOs:72and 73 through a steric construct whereby the C1-C5 bond crosses theplane of the C4-C8 and C3-C7 bonds and is located between them, asillustrated in FIG. 21.

Antibody Ab-A is exemplary and representative of a second group ofantibodies that have a characteristic binding pattern to humansclerostin peptides that is distinct from that obtained for antibodiesAb-C and Ab-D. Ab-A and the group of antibodies it represents bind tothe N22.7-23.5 epitope and have minimal detectable binding to epitopesT19.2, T20, T20.6, T21-22, N14.6 or N18.6, as measured by the ability toblock antibody binding to sclerostin (FIG. 15). A second exemplaryantibody with this binding profile (FIG. 16) is Ab-B. Antibodies havingthis characteristic binding pattern may or may not share amino acidsequence in one or more regions of the antibody molecule. Antibodysimilarity is determined functionally such as by the ability to bind tosclerostin following preincubation with each of the epitopes describedabove. Antibodies that exhibit a binding pattern similar or identical tothat of antibody Ab-A are included in the invention. By “similar to” ismeant, for example, the antibody will exhibit binding to the N22.7-23.5polypeptide whereby this binding will specifically compete out at least50% of the antibody's binding to sclerostin that would otherwise occurin the absence of preincubation with sclerostin or a sclerostin peptide.The antibody will also exhibit little or no detectable binding topolypeptides T19.2, T20, T20.6, T21-22, N14.6 and N18.6, resulting in areduction of 30% or less of the binding that would occur in the absenceof preincubation with sclerostin or a sclerostin peptide.

For example, without being bound by a particular mechanism, the antibodybinding pattern of FIG. 15 suggests that the epitope space to whichantibody Ab-A and other antibodies having the epitope binding pattern ofAb-A bind consists of a polypeptide comprising the Loop 2 region ofsclerostin. Thus, as disclosed herein and with reference to FIG. 19A,the Loop 2 region can be described as a linear peptide, but it acquiresa tertiary structure when it is present in native sclerostin or acystine-knot-containing portion of sclerostin in which the nativedisulfide bond structure is maintained. The linear or tertiary structureof the Loop 2 epitope can affect antibody binding thereto, as discussedin the Examples. A Loop 2 region can comprise the following amino acidsequence: C4GPARLLPNAIGRGKWWRPSGPDFRCS (SEQ ID NO:6). “C4” refers to acysteine residue located at position 86 with reference to SEQ ID NO:1.“C5” refers to a cysteine residue located at position 111 with referenceto SEQ ID NO:1. In native sclerostin protein, C4 is linked to a cysteineat position 144 (C8) by a disulfide bond, and C5 is linked to a cysteineat position 57 (C1) by a disulfide bond. Epitopes derived from the Loop2 region include CGPARLLPNAIGRGKWWRPS (SEQ ID NO:63);GPARLLPNAIGRGKWWRPSG (SEQ ID NO:64); PARLLPNAIGRGKWWRPSGP (SEQ IDNO:65); ARLLPNAIGRGKWWRPSGPD (SEQ ID NO:66); RLLPNAIGRGKWWRPSGPDF (SEQID NO:67); LLPNAIGRGKWWRPSGPDFR (SEQ ID NO:68); and LPNAIGRGKWWRPSGPDFRC(SEQ ID NO:69)

Cross-Blocking Assays

The terms “cross-block”, “cross-blocked” and “cross-blocking” are usedinterchangeably herein to mean the ability of an antibody or otherbinding agent to interfere with the binding of other antibodies orbinding agents to sclerostin.

The extent to which an antibody or other binding agent is able tointerfere with the binding of another to sclerostin, and thereforewhether it can be said to cross-block according to the invention, can bedetermined using competition binding assays. One particularly suitablequantitative assay uses a Biacore machine which can measure the extentof interactions using surface plasmon resonance technology. Anothersuitable quantitative cross-blocking assay uses an ELISA-based approachto measure competition between antibodies or other binding agents interms of their binding to sclerostin.

Biacore Cross-Blocking Assay

The following generally describes a suitable Biacore assay fordetermining whether an antibody or other binding agent cross-blocks oris capable of cross-blocking according to the invention. For conveniencereference is made to two antibodies, but it will be appreciated that theassay can be used with any of the sclerostin binding agents describedherein. The Biacore machine (for example the Biacore 3000) is operatedin line with the manufacturer's recommendations.

Thus in one cross-blocking assay, sclerostin is coupled to a CM5 Biacorechip using standard amine coupling chemistry to generate asclerostin-coated surface. Typically 200-800 resonance units ofsclerostin would be coupled to the chip (an amount that gives easilymeasurable levels of binding but that is readily saturable by theconcentrations of test reagent being used).

The two antibodies (termed A* and B*) to be assessed for their abilityto cross-block each other are mixed at a one to one molar ratio ofbinding sites in a suitable buffer to create the test mixture. Whencalculating the concentrations on a binding site basis the molecularweight of an antibody is assumed to be the total molecular weight of theantibody divided by the number of sclerostin binding sites on thatantibody.

The concentration of each antibody in the test mix should be high enoughto readily saturate the binding sites for that antibody on thesclerostin molecules captured on the Biacore chip. The antibodies in themixture are at the same molar concentration (on a binding basis) andthat concentration would typically be between 1.00 and 1.5 micromolar(on a binding site basis).

Separate solutions containing antibody A* alone and antibody B* aloneare also prepared. Antibody A* and antibody B* in these solutions shouldbe in the same buffer and at the same concentration as in the test mix.

The test mixture is passed over the sclerostin-coated Biacore chip andthe total amount of binding recorded. The chip is then treated in such away as to remove the bound antibodies without damaging the chip-boundsclerostin. Typically this is done by treating the chip with 30 mM HClfor 60 seconds.

The solution of antibody A* alone is then passed over thesclerostin-coated surface and the amount of binding recorded. The chipis again treated to remove all of the bound antibody without damagingthe chip-bound sclerostin.

The solution of antibody B* alone is then passed over thesclerostin-coated surface and the amount of binding recorded.

The maximum theoretical binding of the mixture of antibody A* andantibody B* is next calculated, and is the sum of the binding of eachantibody when passed over the sclerostin surface alone. If the actualrecorded binding of the mixture is less than this theoretical maximumthen the two antibodies are cross-blocking each other.

Thus, in general, a cross-blocking antibody or other binding agentaccording to the invention is one which will bind to sclerostin in theabove Biacore cross-blocking assay such that during the assay and in thepresence of a second antibody or other binding agent of the inventionthe recorded binding is between 80% and 0.1% (e.g. 80% to 4%) of themaximum theoretical binding, specifically between 75% and 0.1% (e.g. 75%to 4%) of the maximum theoretical binding, and more specifically between70% and 0.1% (e.g. 70% to 4%) of maximum theoretical binding (as justdefined above) of the two antibodies or binding agents in combination.

The Biacore assay described above is a primary assay used to determineif antibodies or other binding agents cross-block each other accordingto the invention. On rare occasions particular antibodies or otherbinding agents may not bind to sclerostin coupled via amine chemistry toa CM5 Biacore chip (this usually occurs when the relevant binding siteon sclerostin is masked or destroyed by the coupling to the chip). Insuch cases cross-blocking can be determined using a tagged version ofSclerostin, for example N-terminal His-tagged Sclerostin (R & D Systems,Minneapolis, Minn., USA; 2005 cat #1406-ST-025). In this particularformat, an anti-His antibody would be coupled to the Biacore chip andthen the His-tagged Sclerostin would be passed over the surface of thechip and captured by the anti-His antibody. The cross blocking analysiswould be carried out essentially as described above, except that aftereach chip regeneration cycle, new His-tagged sclerostin would be loadedback onto the anti-His antibody coated surface. In addition to theexample given using N-terminal His-tagged Sclerostin, C-terminalHis-tagged sclerostin could alternatively be used. Furthermore, variousother tags and tag binding protein combinations that are known in theart could be used for such a cross-blocking analysis (e.g. HA tag withanti-HA antibodies; FLAG tag with anti-FLAG antibodies; biotin tag withstreptavidin).

ELISA-Based Cross-Blocking Assay

The following generally describes an ELISA assay for determining whetheran anti-sclerostin antibody or other sclerostin binding agentcross-blocks or is capable of cross-blocking according to the invention.For convenience, reference is made to two antibodies (Ab-X and Ab-Y),but it will be appreciated that the assay can be used with any of thesclerostin binding agents described herein.

The general principal of the assay is to have an anti-sclerostinantibody coated onto the wells of an ELISA plate. An excess amount of asecond, potentially cross-blocking, anti-sclerostin antibody is added insolution (i.e. not bound to the ELISA plate). A limited amount ofsclerostin is then added to the wells. The coated antibody and theantibody in solution compete for binding of the limited number ofsclerostin molecules. The plate is washed to remove sclerostin that hasnot been bound by the coated antibody and to also remove the second,solution phase antibody as well as any complexes formed between thesecond, solution phase antibody and sclerostin. The amount of boundsclerostin is then measured using an appropriate sclerostin detectionreagent. An antibody in solution that is able to cross-block the coatedantibody will be able to cause a decrease in the number of sclerostinmolecules that the coated antibody can bind relative to the number ofsclerostin molecules that the coated antibody can bind in the absence ofthe second, solution phase, antibody.

This assay is described in more detail further below for Ab-X and Ab-Y.In the instance where Ab-X is chosen to be the immobilized antibody, itis coated onto the wells of the ELISA plate, after which the plates areblocked with a suitable blocking solution to minimize non-specificbinding of reagents that are subsequently added. An excess amount ofAb-Y is then added to the ELISA plate such that the moles of Ab-Ysclerostin binding sites per well are at least 10 fold higher than themoles of Ab-X sclerostin binding sites that were used, per well, duringthe coating of the ELISA plate. Sclerostin is then added such that themoles of sclerostin added per well are at least 25-fold lower than themoles of Ab-X sclerostin binding sites that were used for coating eachwell. Following a suitable incubation period the ELISA plate is washedand a sclerostin detection reagent is added to measure the amount ofsclerostin specifically bound by the coated anti-sclerostin antibody (inthis case Ab-X). The background signal for the assay is defined as thesignal obtained in wells with the coated antibody (in this case Ab-X),second solution phase antibody (in this case Ab-Y), sclerostin bufferonly (i.e. no sclerostin) and sclerostin detection reagents. Thepositive control signal for the assay is defined as the signal obtainedin wells with the coated antibody (in this case Ab-X), second solutionphase antibody buffer only (i.e. no second solution phase antibody),sclerostin and sclerostin detection reagents. The ELISA assay needs tobe run in such a manner so as to have the positive control signal be atleast 6 times the background signal.

To avoid any artifacts (e.g. significantly different affinities betweenAb-X and Ab-Y for sclerostin) resulting from the choice of whichantibody to use as the coating antibody and which to use as the second(competitor) antibody, the cross-blocking assay needs to be run in twoformats:

-   -   1) format 1 is where Ab-X is the antibody that is coated onto        the ELISA plate and Ab-Y is the competitor antibody that is in        solution    -   and    -   2) format 2 is where Ab-Y is the antibody that is coated onto        the ELISA plate and Ab-X is the competitor antibody that is in        solution.

Ab-X and Ab-Y are defined as cross-blocking if, either in format 1 or informat 2, the solution phase anti-sclerostin antibody is able to cause areduction of between 60% and 100%, specifically between 70% and 100%,and more specifically between 80% and 100%, of the sclerostin detectionsignal (i.e. the amount of sclerostin bound by the coated antibody) ascompared to the sclerostin detection signal obtained in the absence ofthe solution phase anti-sclerostin antibody (i.e. the positive controlwells).

An example of such an ELISA-based cross blocking assay can be found inExample 7 (“ELISA-based cross-blocking assay”).

Cell Based Neutralization Assay

Mineralization by osteoblast-lineage cells in culture, either primarycells or cell lines, is used as an in vitro model of bone formation.Mineralization takes from about one to six weeks to occur beginning withthe induction of osteoblast-lineage cell differentiation by one or moredifferentiation agents. The overall sequence of events involves cellproliferation, differentiation, extracellular matrix production, matrixmaturation and finally deposition of mineral, which refers tocrystallization and/or deposition of calcium phosphate. This sequence ofevents starting with cell proliferation and differentiation, and endingwith deposition of mineral is referred to herein as mineralization.Measurement of calcium (mineral) is the output of the assay.

MC3T3-E1 cells (Sudo H, Kodama H-A, Amagai Y, Yamamoto S, Kasai S. 1983.In vitro differentiation and calcification in a new clonal osteogeniccell line derived from newborn mouse calvaria. J. Cell Biol. 96:191-198)and subclones of the original cell line can form mineral in culture upongrowth in the presence of differentiating agents. Such subclones includeMC3T3-E1-BF (Smith E, Redman R, Logg C, Coetzee G, Kasahara N, FrenkelB. 2000. Glucocorticoids inhibit developmental stage-specific osteoblastcell cycle. J. Biol. Chem. 275:19992-20001). For both the MC3T3-E1-BFsubclone as well as the original MC3T3-E1 cells, sclerostin can inhibitone or more of the sequence of events leading up to and includingmineral deposition (i.e. sclerostin inhibits mineralization).Anti-sclerostin antibodies that are able to neutralize sclerostin'sinhibitory activity allow for mineralization of the culture in thepresence of sclerostin such that there is a statistically significantincrease in deposition of calcium phosphate (measured as calcium) ascompared to the amount of calcium measured in the sclerostin-only (i.e.no antibody) treatment group. The antibodies used in the cell basedmineralization assay experiments shown in FIGS. 22, 23 and 24 havemolecular weights of about 145 Kd and have 2 sclerostin binding sitesper antibody molecule.

When running the assay with the goal of determining whether a particularanti-sclerostin antibody or anti-sclerostin binding agent can neutralizesclerostin (i.e., is a sclerostin neutralizing antibody or derivativethereof, or is a sclerostin neutralizing binding agent), the amount ofsclerostin used in the assay needs to be the minimum amount ofsclerostin that causes at least a 70%, statistically significant,reduction in deposition of calcium phosphate (measured as calcium) inthe sclerostin-only group, as compared to the amount of calcium measuredin the no sclerostin group. An anti-sclerostin neutralizing antibody oran anti-sclerostin neutralizing binding agent is defined as one thatcauses a statistically significant increase in deposition of calciumphosphate (measured as calcium) as compared to the amount of calciummeasured in the sclerostin-only (i.e. no antibody, no binding agent)treatment group. To determine whether an anti-sclerostin antibody or ananti-sclerostin binding agent is neutralizing or not, the amount ofanti-sclerostin antibody or anti-sclerostin binding agent used in theassay needs to be such that there is an excess of moles of sclerostinbinding sites per well as compared to the number of moles of sclerostinper well. Depending on the potency of the antibody, the fold excess thatmay be required can be 24, 18, 12, 6, 3, or 1.5, and one of skill isfamiliar with the routine practice of testing more than oneconcentration of binding agent. For example, a very potentanti-sclerostin neutralizing antibody or anti-sclerostin neutralizingbinding agent will be able to neutralize sclerostin even when there isless than a 6-fold excess of moles of sclerostin binding sites per wellas compared to the number of moles of sclerostin per well. A less potentanti-sclerostin neutralizing antibody or anti-sclerostin neutralizingbinding agent will be able to neutralize sclerostin only at a 12, 18 or24 fold excess. Sclerostin binding agents within this full range ofpotencies are suitable as neutralizing sclerostin binding agents.Exemplary cell based mineralization assays are described in detail inExample 8.

Anti-sclerostin antibodies and derivatives thereof that can neutralizehuman sclerostin, and sclerostin binding agents that can neutralizehuman sclerostin may be of use in the treatment of humanconditions/disorders that are caused by, associated with, or result inat least one of low bone formation, low bone mineral density, low bonemineral content, low bone mass, low bone quality and low bone strength.

In Vivo Neutralization Assay

Increases in various parameters associated with, or that result from,the stimulation of new bone formation can be measured as an output fromin vivo testing of sclerostin binding agents in order to identify thosebinding agents that are able to neutralize sclerostin and thus able tocause stimulation of new bone formation. Such parameters include variousserum anabolic markers [e.g. osteocalcin, P1NP (n-terminal propeptide oftype 1 procollagen)], histomorphometric markers of bone formation (e.g.osteoblast surface/bone surface; bone formation rate/bone surface;trabecular thickness), bone mineral density, bone mineral content, bonemass, bone quality and bone strength. A sclerostin neutralizing bindingagent is defined as one capable of causing a statistically significantincrease, as compared to vehicle treated animals, in any parameterassociated with, or that results from, the stimulation of new boneformation. Such in vivo testing can be performed in any suitable mammal(e.g. mouse, rat, monkey). An example of such in vivo testing can befound in Example 5 (“In vivo testing of anti-sclerostin monoclonalantibodies”).

Although the amino acid sequence of sclerostin is not 100% identicalacross mammalian species (e.g. mouse sclerostin is not 100% identical tohuman sclerostin), it will be appreciated by one skilled in the art thata sclerostin binding agent that can neutralize, in vivo, the sclerostinof a certain species (e.g. mouse) and that also can bind humansclerostin in vitro is very likely to be able to neutralize humansclerostin in vivo. Thus, such a human sclerostin binding agent (e.g.anti-human sclerostin antibody) may be of use in the treatment of humanconditions/disorders that are caused by, associated with, or result inat least one of low bone formation, low bone mineral density, low bonemineral content, low bone mass, low bone quality and low bone strength.Mice in which homologous recombination had been used to delete the mousesclerostin gene and insert the human sclerostin gene in its place (i.e.human sclerostin gene knock-in mice or human SOST knock-in mice) wouldbe an example of an additional in vivo system.

Pharmaceutical compositions are provided, comprising one of theabove-described binding agents such as at least one of antibody Ab-A,Ab-B, Ab-C, Ab-D and Ab-1 to Ab-24 to human sclerostin, along with apharmaceutically or physiologically acceptable carrier, excipient, ordiluent. Pharmaceutical compositions and methods of treatment aredisclosed in copending application Ser. No. 10/868,497, filed Jun. 16,2004, which claims priority to Ser. No. 60/478,977, both of which areincorporated by reference herein.

The development of suitable dosing and treatment regimens for using theparticular compositions described herein in a variety of treatmentregimens, including e.g., subcutaneous, oral, parenteral, intravenous,intranasal, and intramuscular administration and formulation, is wellknown in the art, some of which are briefly discussed below for generalpurposes of illustration.

In certain applications, the pharmaceutical compositions disclosedherein may be delivered via oral administration to an animal. As such,these compositions may be formulated with an inert diluent or with anassimilable edible carrier, or they may be enclosed in hard- orsoft-shell gelatin capsule, or they may be compressed into tablets, orthey may be incorporated directly with the food of the diet.

In certain circumstances it will be desirable to deliver thepharmaceutical compositions disclosed herein subcutaneously,parenterally, intravenously, intramuscularly, or even intraperitoneally.Such approaches are well known to the skilled artisan, some of which arefurther described, for example, in U.S. Pat. Nos. 5,543,158; 5,641,515and 5,399,363. In certain embodiments, solutions of the active compoundsas free base or pharmacologically acceptable salts may be prepared inwater suitably mixed with a surfactant, such as hydroxypropylcellulose.Dispersions may also be prepared in glycerol, liquid polyethyleneglycols, and mixtures thereof and in oils. Under ordinary conditions ofstorage and use, these preparations generally will contain apreservative to prevent the growth of microorganisms.

Illustrative pharmaceutical forms suitable for injectable use includesterile aqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions (for example, see U.S. Pat. No. 5,466,468). In all cases theform must be sterile and must be fluid to the extent that easysyringability exists. It must be stable under the conditions ofmanufacture and storage and must be preserved against the contaminatingaction of microorganisms, such as bacteria and fungi. The carrier can bea solvent or dispersion medium containing, for example, water, ethanol,polyol (e.g., glycerol, propylene glycol, and liquid polyethyleneglycol, and the like), suitable mixtures thereof, and/or vegetable oils.Proper fluidity may be maintained, for example, by the use of a coating,such as lecithin, by the maintenance of the required particle size inthe case of dispersion and/or by the use of surfactants. The preventionof the action of microorganisms can be facilitated by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars or sodium chloride. Prolonged absorption of the injectablecompositions can be brought about by the use in the compositions ofagents delaying absorption, for example, aluminum monostearate andgelatin.

In one embodiment, for parenteral administration in an aqueous solution,the solution should be suitably buffered if necessary and the liquiddiluent first rendered isotonic with sufficient saline or glucose. Theseparticular aqueous solutions are especially suitable for intravenous,intramuscular, subcutaneous and intraperitoneal administration. In thisconnection, a sterile aqueous medium that can be employed will be knownto those of skill in the art in light of the present disclosure. Forexample, one dosage may be dissolved in 1 ml of isotonic NaCl solutionand either added to 1000 ml of hypodermoclysis fluid or injected at theproposed site of infusion, (see for example, Remington's PharmaceuticalSciences, 15th ed., pp. 1035-1038 and 1570-1580). Some variation indosage will necessarily occur depending on the condition of the subjectbeing treated. Moreover, for human administration, preparations will ofcourse preferably meet sterility, pyrogenicity, and the general safetyand purity standards as required by FDA Office of Biologics standards.

In another embodiment of the invention, the compositions disclosedherein may be formulated in a neutral or salt form. Illustrativepharmaceutically-acceptable salts include the acid addition salts(formed with the free amino groups of the protein) and which are formedwith inorganic acids such as, for example, hydrochloric or phosphoricacids, or such organic acids as acetic, oxalic, tartaric, mandelic, andthe like. Salts formed with the free carboxyl groups can also be derivedfrom inorganic bases such as, for example, sodium, potassium, ammonium,calcium, or ferric hydroxides, and such organic bases as isopropylamine,trimethylamine, histidine, procaine and the like. Upon formulation,solutions will be administered in a manner compatible with the dosageformulation and in such amount as is therapeutically effective.

The carriers can further comprise any and all solvents, dispersionmedia, vehicles, coatings, diluents, antibacterial and antifungalagents, isotonic and absorption delaying agents, buffers, carriersolutions, suspensions, colloids, and the like. The use of such mediaand agents for pharmaceutical active substances is well known in theart. Except insofar as any conventional media or agent is incompatiblewith the active ingredient, its use in the therapeutic compositions iscontemplated. Supplementary active ingredients can also be incorporatedinto the compositions. The phrase “pharmaceutically-acceptable” refersto molecular entities and compositions that do not produce an allergicor similar untoward reaction when administered to a human.

In certain embodiments, liposomes, nanocapsules, microparticles, lipidparticles, vesicles, and the like, are used for the introduction of thecompositions of the present invention into suitable hostcells/organisms. In particular, the compositions of the presentinvention may be formulated for delivery either encapsulated in a lipidparticle, a liposome, a vesicle, a nanosphere, or a nanoparticle or thelike. Alternatively, compositions of the present invention can be bound,either covalently or non-covalently, to the surface of such carriervehicles.

The formation and use of liposome and liposome-like preparations aspotential drug carriers is generally known to those of skill in the art(see for example, Lasic, Trends Biotechnol. 16(7):307-21, 1998;Takakura, Nippon Rinsho 56(3):691-95, 1998; Chandran et al., Indian J.Exp. Biol. 35(8):801-09, 1997; Margalit, Crit. Rev. Ther. Drug CarrierSyst. 12(2-3):233-61, 1995; U.S. Pat. Nos. 5,567,434; 5,552,157;5,565,213; 5,738,868 and 5,795,587, each specifically incorporatedherein by reference in its entirety). The use of liposomes does notappear to be associated with autoimmune responses or unacceptabletoxicity after systemic delivery. In certain embodiments, liposomes areformed from phospholipids that are dispersed in an aqueous medium andspontaneously form multilamellar concentric bilayer vesicles (alsotermed multilamellar vesicles (MLVs)).

Alternatively, in other embodiments, the invention provides forpharmaceutically-acceptable nanocapsule formulations of the compositionsof the present invention. Nanocapsules can generally entrap compounds ina stable and reproducible way (see, for example, Quintanar-Guerrero etal., Drug Dev. Ind. Pharm. 24(12):1113-28, 1998). To avoid side effectsdue to intracellular polymeric overloading, such ultrafine particles(sized around 0.1 μm) may be designed using polymers able to be degradedin vivo. Such particles can be made as described, for example, byCouvreur et al., Crit. Rev. Ther. Drug Carrier Syst. 5(1):1-20, 1988;zur Muhlen et al., Eur. I Pharm. Biopharm. 45(2):149-55, 1998; Zambauxet al., J. Controlled Release 50(1-3):31-40, 1998; and U.S. Pat. No.5,145,684.

In addition, pharmaceutical compositions of the present invention may beplaced within containers, along with packaging material that providesinstructions regarding the use of such pharmaceutical compositions.Generally, such instructions will include a tangible expressiondescribing the reagent concentration, as well as within certainembodiments, relative amounts of excipient ingredients or diluents(e.g., water, saline or PBS) that may be necessary to reconstitute thepharmaceutical composition.

The dose administered may range from 0.01 mg/kg to 100 mg/kg of bodyweight. As will be evident to one of skill in the art, the amount andfrequency of administration will depend, of course, on such factors asthe nature and severity of the indication being treated, the desiredresponse, the condition of the patient, and so forth. Typically, thecompositions may be administered by a variety of techniques, as notedabove.

Increases in bone mineral content and/or bone mineral density may bedetermined directly through the use of X-rays (e.g., Dual Energy X-rayAbsorptometry or “DEXA”), or by inference through the measurement of 1)markers of bone formation and/or osteoblast activity, such as, but notlimited to, osteoblast specific alkaline phosphatase, osteocalcin, type1 procollagen C′ propeptide (PICP), total alkaline phosphatase (seeCornier, Curr. Opin. in Rheu. 7:243(1995)) and serum procollagen 1N-terminal propeptide (P1NP) and/or 2) markers of bone resorption and/orosteoclast activity including, but not limited to, pyridinoline,deoxypryridinoline, N-telopeptide, urinary hydroxyproline, plasmatartrate-resistant acid phosphatases, and galactosyl hydroxylysine; (seeCornier, id), serum TRAP 5b (tartrate-resistant acid phosphatase isoform5b) and serum cross-linked C-telopeptide (sCTXI). The amount of bonemass may also be calculated from body weights or by using other methods(see Guinness-Hey, Metab. Bone Dis. Relat. Res. 5:177-181, 1984).Animals and particular animal models are used in the art for testing theeffect of the compositions and methods of the invention on, for example,parameters of bone loss, bone resorption, bone formation, bone strengthor bone mineralization that mimic conditions of human disease such asosteoporosis and osteopenias. Examples of such models include theovariectomized rat model (Kalu, D. N., The ovariectomized rat model ofpostmenopausal bone loss. Bone and Mineral 15:175-192 (1991); Frost, H.M. and Jee, W. S. S. On the rat model of human osteopenias andosteoporosis. Bone and Mineral 18:227-236 (1992); and Jee, W. S. S. andYao, W., Overview: animal models of osteopenia and osteoporosis. J.Musculoskel. Neuron. Interact. 1:193-207 (2001)).

Particular conditions which may be treated by the compositions of thepresent invention include dysplasias, wherein growth or development ofbone is abnormal and a wide variety of causes of osteopenia,osteoporosis and bone loss. Representative examples of such conditionsinclude achondroplasia, cleidocranial dysostosis, enchondromatosis,fibrous dysplasia, Gaucher's Disease, hypophosphatemic rickets, Marfan'ssyndrome, multiple hereditary exotoses, neurofibromatosis, osteogenesisimperfecta, osteopetrosis, osteopoikilosis, sclerotic lesions,pseudoarthrosis, and pyogenic osteomyelitis, periodontal disease,anti-epileptic drug induced bone loss, primary and secondaryhyperparathyroidism, familial hyperparathyroidism syndromes,weightlessness induced bone loss, osteoporosis in men, postmenopausalbone loss, osteoarthritis, renal osteodystrophy, infiltrative disordersof bone, oral bone loss, osteonecrosis of the jaw, juvenile Paget'sdisease, melorheostosis, metabolic bone diseases, mastocytosis, sicklecell anemia/disease, organ transplant related bone loss, kidneytransplant related bone loss, systemic lupus erythematosus, ankylosingspondylitis, epilepsy, juvenile arthritides, thalassemia,mucopolysaccharidoses, fabry disease, turner syndrome, Down Syndrome,Klinefelter Syndrome, leprosy, Perthes' Disease, adolescent idiopathicscoliosis, infantile onset multi-system inflammatory disease, WinchesterSyndrome, Menkes Disease, Wilson's Disease, ischemic bone disease (suchas Legg-Calve-Perthes disease, regional migratory osteoporosis), anemicstates, conditions caused by steroids, glucocorticoid-induced bone loss,heparin-induced bone loss, bone marrow disorders, scurvy, malnutrition,calcium deficiency, idiopathic osteopenia or osteoporosis, congenitalosteopenia or osteoporosis, alcoholism, chronic liver disease,postmenopausal state, chronic inflammatory conditions, rheumatoidarthritis, inflammatory bowel disease, ulcerative colitis, inflammatorycolitis, Crohn's disease, oligomenorrhea, amenorrhea, pregnancy,diabetes mellitus, hyperthyroidism, thyroid disorders, parathyroiddisorders, Cushing's disease, acromegaly, hypogonadism, immobilizationor disuse, reflex sympathetic dystrophy syndrome, regional osteoporosis,osteomalacia, bone loss associated with joint replacement, HIVassociated bone loss, bone loss associated with loss of growth hormone,bone loss associated with cystic fibrosis, fibrous dysplasia,chemotherapy associated bone loss, tumor induced bone loss,cancer-related bone loss, hormone ablative bone loss, multiple myeloma,drug-induced bone loss, anorexia nervosa, disease associated facial boneloss, disease associated cranial bone loss, disease associated bone lossof the jaw, disease associated bone loss of the skull, and bone lossassociated with space travel. Further conditions relate to bone lossassociated with aging, including facial bone loss associated with aging,cranial bone loss associated with aging, jaw bone loss associated withaging, and skull bone loss associated with aging.

Compositions of the present invention may also be useful for improvingoutcomes in orthopedic procedures, dental procedures, implant surgery,joint replacement, bone grafting, bone cosmetic surgery and bone repairsuch as fracture healing, nonunion healing, delayed union healing andfacial reconstruction. One or more compositions may be administeredbefore, during and/or after the procedure, replacement, graft, surgeryor repair.

The invention also provides a diagnostic kit comprising at least oneanti-sclerostin binding agent according to the present invention. Thebinding agent may be an antibody. In addition, such a kit may optionallycomprise one or more of the following:

-   -   (1) instructions for using the one or more binding agent(s) for        screening, diagnosis, prognosis, therapeutic monitoring or any        combination of these applications;    -   (2) a labeled binding partner to the anti-sclerostin binding        agent(s);    -   (3) a solid phase (such as a reagent strip) upon which the        anti-sclerostin binding agent(s) is immobilized; and    -   (4) a label or insert indicating regulatory approval for        screening, diagnostic, prognostic or therapeutic use or any        combination thereof.        If no labeled binding partner to the binding agent(s) is        provided, the binding agent(s) itself can be labeled with one or        more of a detectable marker(s), e.g. a chemiluminescent,        enzymatic, fluorescent, or radioactive moiety.

The following examples are offered by way of illustration, and not byway of limitation.

EXAMPLES Example 1 Recombinant Expression of Sclerostin

Recombinant human sclerostin/SOST is commercially available from R&DSystems (Minneapolis, Minn., USA; 2006 cat #1406-ST-025). Additionally,recombinant mouse sclerostin/SOST is commercially available from R&DSystems (Minneapolis, Minn., USA; 2006 cat #1589-ST-025).

Alternatively, the different species of sclerostin can be expressedtransiently in serum-free suspension adapted 293T or 293EBNA cells.Transfections can be performed as 500 mL or 1 L cultures. The followingreagents and materials are available from Gibco BRL (now Invitrogen,Carlsbad, Calif.). Catalog numbers are listed in parentheses: serum-freeDMEM (21068-028); DMEM/F12 (3:1) (21068/11765); 1×Insulin-Transferrin-Selenium Supplement (51500-056); 1× Pen Strep Glut(10378-016); 2 mM 1-Glutamine (25030-081); 20 mM HEPES (15630-080);0.01% Pluronic F68 (24040-032). Briefly, the cell inoculum (5.0-10.0×10⁵cells/mL×culture volume) is centrifuged at 2,500 RPM for 10 minutes at4° C. to remove the conditioned medium.

The cells are resuspended in serum-free DMEM and centrifuged again at2,500 RPM for 10 minutes at 4° C. After aspirating the wash solution,the cells are resuspended in growth medium [DMEM/F12 (3:1)+1×Insulin-Transferrin-Selenium Supplement+1× Pen Strep Glut+2 mML-Glutamine+20 mM HEPES+0.01% Pluronic F68] in a 1 L or 3 L spinnerflask culture. The spinner flask culture is maintained on magnetic stirplate at 125 RPM which is placed in a humidified incubator maintained at37° C. and 5% CO₂. The mammalian expression plasmid DNA (e.g. pcDNA3.1,pCEP4, Invitrogen Life Technologies, Carlsbad, Calif.), containing thecomplete coding region (and stop codon) of sclerostin with a Kozakconsensus sequence (e.g., CCACC) directly 5′ of the start site ATG, iscomplexed to the transfection reagent in a 50 mL conical tube.

The DNA-transfection reagent complex can be prepared in 5-10% of thefinal culture volume in serum-free DMEM or OPTI-MEM. The transfectionreagents that can be used for this purpose include X-tremeGene RO-1539(Roche Applied Science, Indianapolis, Ind.), FuGene6 (Roche AppliedScience, Indianapolis, Ind.), Lipofectamine 2000 (Invitrogen, Carlsbad,Calif.) and 293fectin (Invitrogen, Carlsbad, Calif.). 1-5 μg plasmidDNA/mL culture is first added to serum-free DMEM, followed by 1-5 μltransfection reagent/mL culture. The complexes can be incubated at roomtemperature for approximately 10-30 minutes and then added to the cellsin the spinner flask. The transfection/expression can be performed for4-7 days, after which the conditioned medium (CM) is harvested bycentrifugation at 4,000 RPM for 60 minutes at 4° C.

Example 2 Purification of Recombinant Sclerostin

Recombinant sclerostin was purified from mammalian host cells asfollows. All purification processes were carried out at roomtemperature. One purification scheme was used to purify various speciesof sclerostin, including murine and human sclerostin. The purificationscheme used affinity chromatography followed by cation exchangechromatography.

Heparin Chromatography

The mammalian host cell conditioned medium (CM) was centrifuged in aBeckman J6-M1 centrifuge at 4000 rpm for 1 hour at 4° C. to remove celldebris. The CM supernatant was then filtered through a sterile 0.2 μmfilter. (At this point the sterile filtered CM may be optionally storedfrozen until purification.) If the CM was frozen, it was thawed at thefollowing temperatures, or combination thereof: 4° C., room temperatureor warm water. Following thawing the CM was filtered through a sterile0.2 μm filter and optionally concentrated by tangential flowultrafiltration (TFF) using a 10 kD molecular weight cut-off membrane.The CM concentrate was filtered through a sterile 0.2 μm filter and thenloaded onto a Heparin High Performance (Heparin HP) column (GEHealthcare, formerly Amersham Biosciences) equilibrated in PBS.Alternatively, the filtered CM supernatant may be loaded directly ontothe Heparin HP column equilibrated in PBS.

After loading, the Heparin HP column was washed with PBS until theabsorbance at 280 nm of the flow-through returned to baseline (i.e.,absorbance measured before loading CM supernatant). The sclerostin wasthen eluted from the column using a linear gradient from 150 mM to 2Msodium chloride in PBS. The absorbance at 280 nm of the eluate wasmonitored and fractions containing protein were collected. The fractionswere then assayed by Coomassie-stained SDS-PAGE to identify fractionscontaining a polypeptide that migrates at the size of glycosylatedsclerostin. The appropriate fractions from the column were combined tomake the Heparin HP pool.

Cation Exchange Chromatography

The sclerostin eluted from the Heparin HP column was further purified bycation exchange chromatography using SP High Performance (SPHP)chromatography media (GE Healthcare, formerly Amersham Biosciences). TheHeparin HP pool was buffer exchanged into PBS by dialysis using 10,000MWCO membranes (Pierce Slide-A-Lyzer). The dialyzed Heparin HP pool wasthen loaded onto an SPHP column equilibrated in PBS. After loading, thecolumn was washed with PBS until the absorbance at 280 nm of theflow-through returned to baseline. The sclerostin was then eluted fromthe SPHP column using a linear gradient from 150 mM to 1 M sodiumchloride in PBS. The absorbance at 280 nm of the eluate was monitoredand the eluted sclerostin was collected in fractions. The fractions werethen assayed by Coomassie-stained SDS-PAGE to identify fractionscontaining a polypeptide that migrates at the size of glycosylatedsclerostin. The appropriate fractions from the column were combined tomake the SPHP pool.

Formulation

Following purification, the SPHP pool was formulated in PBS by dialysisusing 10,000 MWCO membranes (Pierce Slide-A-Lyzer). If concentration ofsclerostin was necessary, a centrifugal device (Amicon Centricon orCentriprep) with a 10,000 MWCO membrane was used. Following formulationthe sclerostin was filtered through a sterile 0.2 μm filter and storedat 4° C. or frozen.

Example 3 Peptide Binding ELISA

A series of overlapping peptides (each peptide being approximately 20-25amino acids long) were synthesized based on the known amino acidsequence of rat sclerostin (SEQ ID NO:98). The peptides were designedsuch that they all contained a reduced cysteine residue; an additionalcysteine was included at the C-terminus of each peptide which did notalready contain one in its sequence. This enabled the peptides to bebound to the assay plates by covalent coupling, using commerciallyavailable sulfhydryl binding plates (Costar), at a concentration of 1μg/ml, in phosphate buffered saline (PBS: pH 6.5) containing 1 mM EDTA.Following incubation for 1 hour at room temperature, the plates werewashed three times with PBS containing 0.5% Tween 20. The plates wereblocked by incubation with a PBS solution containing 0.5% fish skingelatin (Sigma) for 30 minutes at room temperature and then washed threetimes in PBS containing 0.5% Tween 20.

Antibodies to be tested were diluted to 1 μg/ml in PBS containing 0.5%fish skin gelatin and incubated with the peptide-coated plates for 1hour at room temperature. Excess antibody was removed by three washeswith PBS, 0.5% Tween 20. The plates were then incubated with anappropriate secondary antibody conjugated to horseradish peroxidase(diluted appropriately in PBS containing 0.5% Tween 20) and capable ofbinding to the antibody of interest. The plates were then washed threetimes: once with PBS containing 0.5% Tween 20, and twice with PBS.Finally the plates were incubated with a horseradish peroxidasechromogenic substrate (TMB-Stable Stop, RDI) for 5 minutes at roomtemperature, the color development was stopped with acid, and theplates' optical density measured at 450 nm.

Materials

-   -   Costar's Sulfhydryl Binding Plates (VWR #29442-278)    -   Coating Buffer: 1×PBS PH 6.5+1 mM EDTA    -   Blocking Buffer: 1×PBS+0.5% Fish Skin Gelatin (PBS from CS; FSG        from Sigma # G 7765)    -   Wash Buffer: 1×PBS+0.5% Tween 20    -   Rat Sclerostin peptides    -   Antibody Samples: Transient Ab, Purified recombinant Ab, rabbit        Serum, etc.    -   Appropriate secondary Ab: Goat-anti-Rabbit/Mouse-HRP (Jackson        Immuno Research, 115-036-072)    -   TMB-Stable Stop (RDI # RDI-TMBSX-1 L)    -   0.5M HCl

Methods were as Follows:

-   -   1. Coat plates with 100 μl/well of rat sclerostin peptide        diluted in 1×PBS PH 6.5+EDTA at 1 μg/ml. Incubate plates 1 hour        at room temperature. (Plates should be used within 30 minutes of        opening).    -   2. Wash plates 3× with wash buffer.    -   3. Block plates with 200 ul/well blocking buffer. Incubate        plates 30 minutes at room temp.    -   4. Repeat washing as described in (2).    -   5. Incubate plates with 50 ul/well of samples diluted in        blocking buffer—Serum titers starting at 1:100; Transient        Recombinant Ab use neat; Purified recombinant Ab use at 1 μg/ml        (all samples run in duplicates). Incubate plates 1 h at room        temp.    -   6. Wash plates as described in (2).    -   7. Incubate plates with 50 μl/well of appropriate Secondary        Antibody (HRP labeled) diluted 1:1600 in Blocking Buffer.        Incubate plates 1 hour at room temperature.    -   8. Wash plates 1× wash buffer, 2×PBS    -   9. Incubate plates with 50 μl/well of TMB, 5 minutes at room        temp.    -   10. Stop reaction with 50 μl/well 0.5M HCl.    -   11. Read plates at 450 nm wavelength.

The following peptides sequences were screened as described above:

(SEQ ID NO: 82) QGWQAFKNDATEIIPGLREYPEPP (SEQ ID NO: 83)TEIIPGLREYPEPPQELENN (SEQ ID NO: 84) PEPPQELENNQTMNRAENGG (SEQ ID NO:85) ENGGRPPHHPYDTKDVSEYS (SEQ ID NO: 86) CRELHYTRFVTDGP (SEQ ID NO: 87)CRELHYTRFVTDGPSRSAKPVTELV (SEQ ID NO: 88) CRSAKPVTELVSSGQSGPRARLL (SEQID NO: 89) CGPARLLPNAIGRVKWWRPNGPDFR (SEQ ID NO: 90)RAQRVQLLCPGGAAPRSRKV (SEQ ID NO: 91) PGGAAPRSRKVRLVAS (SEQ ID NO: 92)KRLTRFHNQSELKDFGPETARPQ (SEQ ID NO: 93) IPDRYAQRVQLLSPGG (SEQ ID NO: 94)SELKDFGPETARPQKGRKPRPRAR (SEQ ID NO: 95) KGRKPRPRARGAKANQAELENAY (SEQ IDNO: 96) PNAIGRVKWWRPNGPDFR (SEQ ID NO: 97) KWWRPNGPDFRCIPDRYRAQRV.

A high-affinity neutralizing antibody (Ab-19) bound to two overlappingpeptide sequences:

(SEQ ID NO: 96) PNAIGRVKWWRPNGPDFR and (SEQ ID NO: 97)KWWRPNGPDFRCIPDRYRAQRV.

This procedure allows the recognition of epitopes for antibodies thatreact with apparent linear epitopes. Peptides that contain all or partof the antibody binding site will bind antibody and thus be detected.

Example 4 Identification of Human Sclerostin Epitopes SclerostinStructure

Mature form (signal peptide removed) human sclerostin is a 190 aminoacid protein (FIG. 8). FIG. 9 shows a schematic of the general structureof sclerostin with an N-terminal arm (from the N-terminal Q toCysteine1) and a C-terminal arm (from Cysteine8 to the terminal Y).Sandwiched in between these two arms there is the cystine-knot structureand three loops which are designated Loop1, Loop2 and Loop 3. The fourdisulfide bonds in sclerostin are Cys1 at sequence position 57 linked toCys5 at sequence position 111 (referred to as C1-C5), Cyst at sequenceposition 71 linked to Cys6 at sequence position 125 (referred to asC2-C6), Cys3 at sequence position 82 linked to Cys7 at sequence position142 (referred to as C3-C7), Cys4 at sequence position 86 linked to Cys8at sequence position 144 (referred to as C4-C8). The eight-membered ringstructure is formed via C3-C7 and C4-C8 disulfide bonding. This ringstructure, together with the C1-C5 disulfide bond penetrating throughthe ring, forms a typical cystine-knot. C2-C6, which is not part of thecystine-knot, brings two large loop structures, loop 1 (residues 57 to82) and loop 3 (residues 111 to 142) close together. Loop 2 goes from C4(residue 86) to C5 (residue 111).

Experimental

The general approach for characterizing the epitopes bound byanti-sclerostin monoclonal antibodies involved fragmenting humanSclerostin into peptides with different proteases, determining thesequence of the various human sclerostin peptides, isolating thesepeptides and testing each of them for their ability to bind to aparticular monoclonal antibody using a Biacore-based “human sclerostinpeptide epitope competition binding assay.”. The resulting datapermitted the location of the binding epitope to be determined.

The peptide digests were subjected to HPLC peptide mapping; theindividual peaks were collected, and the peptides identified and mappedby matrix assisted laser desorption mass spectrometry (MALDI-MS) andelectrospray ionization LC-MS (ESI-LC-MS) analyses and/or by N-terminalsequencing. All HPLC analyses for these studies were performed using areverse-phase C8 column (2.1 mm i.d.×15 cm length). HPLC peptide mappingwas performed with a linear gradient from 0.05% trifloroacetic acid(mobile phase A) to 90% acetonitrile in 0.05% trifuoroacetic acid.Columns were developed over 50 minutes at a flow rate of 0.2 ml/min.

Trypsin and AspN Endoproteinase Digestions

Mature form human sclerostin was digested with trypsin, which cleavesafter arginine and lysine, or with AspN. About 200 μg of sclerostin at0.5-1.0 mg/ml was incubated in PBS (pH 7.2) for 20 hrs at 37° C. with 8μg of either trypsin or AspN.

Trypsin Digestion

HPLC chromatography of the trypsin digests yielded several major peaks(FIG. 10A). Sequence analysis was conducted on the peptide peaksrecovered from HPLC after trypsin digestion. On-line ESI LC-MS analysisof the peptide digest was also performed to determine the precise massof the peptides that were separated by HPLC. The identity of thepeptides present in the peptide peaks was thus determined (FIG. 11).FIG. 13 shows the alignment of various peptide sequences (T19.2, T20,T20.6, T21-22) along the sclerostin sequence. The number following eachT (e.g., T19.2) reflects the retention time. T19.2 contains two peptides(one from loop 1 and one from loop 3) linked by the C2-C6 disulfidebond. T20 contains two peptides held together by the cystine-knotstructure, with intact loops 1 and 3 held together by the C2-C6disulfide and with most of loop 2 absent. T20.6 contains four sequencesheld together by the cystine-knot structure, but is missing part of loop1 and 3 (the T19.2 part) and is missing most of loop 2. T21-22 is almostidentical to T20 but has 3 additional amino acids in the loop 2 region.

AspN Digestion

HPLC chromatography of the AspN digests yielded several major peaks(FIG. 10B). Sequence analysis was conducted on the peptide peaksrecovered from HPLC. On-line ESI LC-MS analysis of the peptide digestwas also performed to determine the precise mass of the peptides thatwere separated by HPLC. The identity of the peptides present in thepeptide peaks from the AspN digestion was thus determined (FIG. 12).FIG. 14 shows the alignment of various peptide sequences (AspN14.6,AspN18.6, AspN22.7-23.5) along the sclerostin sequence. The numberfollowing each AspN (e.g. AspN18.6) reflects the retention time.AspN14.6 contains three short peptides from both the N- and C-terminalarms of sclerostin, while AspN18.6 is a larger peptide from theN-terminal arm of sclerostin. AspN22.7-23.5 contains a single peptidefragment of 104 amino acids the encompasses all eight cysteines (thefour disulfide bonds), the cystine-knot and all of loops 1, 2 and 3.

The strategy for characterizing the epitopes was to use these varioustrypsin and AspN generated human sclerostin peptides and determine whichpeptides could still be bound by the various Antibodies (Ab-A, Ab-B,Ab-C and Ab-D). Specifically this was tested in a Biacore-based “humansclerostin peptide epitope competition binding assay” where the bindingof a particular monoclonal antibody to human sclerostin immobilized onthe Biacore chip was determine in the presence or absence of each of thevarious isolated trypsin and AspN HPLC peptide fractions. In the absenceof any competing peptides, the particular monoclonal antibody was ableto bind the human sclerostin on the chip and produce a resonance unit,RU, response. Preincubation of the particular monoclonal antibody withintact human sclerostin in solution, followed by testing of binding tothe chip, demonstrated that the binding of the Mab to human sclerostinin solution prevented the binding of the Mab to the human sclerostin onthe chip, thus validating the general principal of this competitionassay.

This general procedure was repeated individually for each peptide. Arobust RU response was taken to indicate that the particular peptidebeing tested could not bind the Mab in solution (hence the Mab was freeto bind the human sclerostin that had been immobilized on the chip).Conversely, the absence of a robust RU response indicated that the Mabwas able to bind the sclerostin peptide in solution. These bindingpatterns, couple with the known identity of the various sclerostinpeptides, were used to determine the epitopes of sclerostin that werebound by anti-sclerostin antibodies Ab-A, Ab-B, Ab-C and Ab-D.

Biacore-Based Human Sclerostin Peptide Epitope Competition Binding AssayPreparation of Human Sclerostin Surface:

Immobilization of mature form human sclerostin to a BIAcore sensor chip(CM5) surface was performed according to manufacturer's instructions.Briefly, carboxyl groups on the sensor chip surfaces were activated byinjecting 60 μL of a mixture containing 0.2 MN-ethyl-N′-(dimethylaminopropyl) carbodiimide (EDC) and 0.05 MN-hydroxysuccinimide (NHS). Human sclerostin was diluted in 10 mM sodiumacetate, pH 4.0 at a concentration of 20 μg/mL followed by injectingover the activated CM5 surface. Excess reactive groups on the surfaceswere deactivated by injecting 60 μL of 1 M ethanolamine. Finalimmobilized levels were ˜5000 resonance units (RU) for the humansclerostin surface. A blank, mock-coupled reference surface was alsoprepared on the sensor chips.

Binding Specificity Analysis:

1× Phosphate-buffered saline without calcium chloride or magnesiumchloride was from Gibco/Invitrogen, Carlsbad, Calif. Bovine serumalbumin, fraction V, IgG-free was from Sigma-Aldrich, St. Louis, Mo.Each Mab (2 nM) was separately incubated with 20 nM human sclerostin ora particular human sclerostin peptide (note: there are 3 unlinkedpeptides in AspN14.6) in sample buffer (1×PBS+0.005% P-20+0.1 mg/mL BSA)before injection over the immobilized human sclerostin surface. The flowrate for sample injection was 5 μL/min followed by surface regenerationusing 1 M NaCl in 8 mM Glycine, pH 2.0 at 30 μL/min for 30 seconds. Thedata was analyzed using BIAevaluation 3.2, and is presented in FIG. 15(Ab-A), FIG. 16 (Ab-B), FIG. 17 (Ab-C) and FIG. 18 (Ab-D).

Loop 2 and T20.6 Epitopes:

The sclerostin peptide binding pattern for two representative antibodies(Ab-A and Ab-B) were virtually identical (FIG. 15 and FIG. 16) andshowed that both of these Antibodies could only bind the AspN22.7-23.5peptide. The unique difference between AspN22.7-23.5 and all the othersclerostin peptides is that AspN22.7-23.5 contains an intact loop 2.This shows that Ab-A and Ab-B bind the loop 2 region of sclerostin thusdefining the loop 2 epitope (FIG. 19A). The sclerostin peptide bindingpattern for Ab-C and Ab-D were virtually identical to each other (FIG.17 and FIG. 18) but completely distinct from that found for Ab-A andAb-B. Of the peptides tested in this Example, the most diminutivepeptide that Ab-C and Ab-D could bind to was the T20.6 peptide. Thisresult defines the T20.6 epitope (FIG. 19B).

Protease Protection Assay:

The general principle of this assay is that binding of a Mab tosclerostin can result in protection of certain specific proteasecleavage sites and this information can be used to determine the regionof sclerostin to where the Mab binds.

“T20.6 Derivative 1 (Cystine-Knot+4 Arms)” Epitope:

FIG. 20 shows the HPLC peptide maps for a human sclerostin Ab-D complex(FIG. 20A: human sclerostin was preincubated at a 1:1 molar ratio withAb-D prior to digestion with trypsin as described above) and humansclerostin alone (FIG. 20B: human sclerostin was digested with trypsinas described above). The peptide peaks of T19.2 and T20.6 in FIG. 20Ashowed a clear reduction in their respective peak height, as compared toFIG. 20B. This reduction in peak heights was accompanied by an increasein peak height for peptides T20 and T21-22. These data indicate thatbasic amino acid residues in loop 1 and loop 3, which in the absence ofAb-D were cleaved by trypsin to generate peptides T19.2 and T20.6, wereresistant to cleavage by trypsin when Ab-D was prebound to sclerostin.The presence of T20, T20.6 and T21-22 indicates that loop 2 was stillcleaved efficiently when Ab-D was prebound to sclerostin. These dataindicate that Ab-D bound on the loop 1 and loop 3 side of the T20.6epitope thus defining the smaller “T20.6 derivative 1 (cystine-knot+4arms)” epitope shown in FIG. 21.

Example 5 In Vivo Testing of Anti-Sclerostin Monoclonal Antibodies inMice

Four week-old BDF1 male mice were obtained from Charles RiverLaboratories (Raleigh, N.C.) and housed in clean caging, five animalsper cage. Room temperature was maintained between 68 and 72° F., andrelative humidity was maintained between 34 and 73%. The laboratoryhousing the cages had a 12-hour light/dark cycle and met all AAALACspecifications. Clinical observations of all mice on study occurred oncedaily.

Purified anti-sclerostin monoclonal antibodies (Ab-A FIG. 1; Ab-B FIG.2; Ab-C FIG. 3; Ab-D FIG. 4) were diluted in sterile Dulbecco'sphosphate buffered saline. Mice were injected with anti-sclerostinAntibodies or PBS vehicle subcutaneously at 21 μl per gram body weight,two times per week (Monday and Thursday) at 25 mg/kg. Human PTH (1-34)was diluted in PTH buffer (0.001 N HCl, 0.15 M NaCl, 2% BSA), and dosedsubcutaneously at 21 μl per gram body weight five times per week(Monday, Tuesday, Wednesday, Thursday, Friday) at 100 μg/kg as apositive control (FIGS. 5 and 6). Number of mice per group was N=5 inFIGS. 5 and 6, and N=6 in FIG. 7.

PIXImus In Vivo Bone Densitometry

Bone mineral density (BMD) was determined weekly at the proximal tibialmetaphysis and lumbar vertebrae by peripheral Dual Energy X-rayAbsorptometry (pDEXA) with the PIXImus2 system from GE/Lunar MedicalSystems, Madison, Wis. A 25 mm² region of interest (ROI) was placed toinclude the proximal articular surface, the epiphysis, and the proximalend on the metaphysis of the tibia. A region of interest (ROI) wasplaced to include the lumbar vertebrae (L1-L5). The proximal tibia andlumbar regions were analyzed to determine total bone mineral density.Group means were reported±Standard Deviation and compared to the vehicletreatment group for statistical analysis.

Statistical Analysis

Statistical analysis was performed with a Dunnett's and Tukey-Kramer(using MS Excel and JMP v. 5.0. for the BMD data). Group means for eachdata set were considered significantly different when the P value wasless than 0.05 (P<0.05).

Sclerostin Neutralizing Activity of Antibodies

The statistically significant increases in BMD as compared to vehicleseen for each of Ab-A (FIG. 5), Ab-B (FIG. 5), Ab-C (FIG. 6) and Ab-D(FIG. 7) demonstrates that these four antibodies are sclerostinneutralizing antibodies. Furthermore this data shows that, foranti-sclerostin antibodies that bind mouse sclerostin, treatment andanalysis of mice as described above can be used to identify sclerostinneutralizing antibodies.

Example 6 Screening Assay for Antibodies that Block Binding of anAntibody to Human Sclerostin

Human sclerostin was coupled to a CM5 Biacore chip using standard aminecoupling chemistry to generate a sclerostin coated surface. 300resonance units of sclerostin were coupled to the surface.

The antibodies to be tested were diluted to a concentration of 200 ug/mlin HBS-EP buffer (being 10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA,0.005% (v/v) Surfactant P20) and then mixed in a one to one molar ratio(on a binding site basis) to generate the test mixture. This testmixture thus contained each antibody at a concentration of 100 ug/ml(1.3 um on a binding site basis). Separate solutions containing each ofthe antibodies in the test mix alone were also prepared. These solutionscontained the individual antibodies in HBS-EP buffer at a concentrationof 100 ug/ml (1.3 um on a binding site basis).

20 μL of the test mixture was passed over the sclerostin-coated chip ata flow rate of 10 μL/min and the amount of binding recorded. The chipwas then treated with two 60 second pulses of 30 mM HCl to remove all ofthe bound antibody. A solution containing only one of the antibodies ofthe test mixture (at 1.3 μM in the same buffer as the test mixture on abinding site basis) was then passed over the chip in the same manner asthe test mixture and the amount of binding recorded. The chip was againtreated to remove all of the bound antibody and finally a solutioncontaining the other antibody from the test mixture alone (at 1.3 μM inthe same buffer as the test mixture on a binding site basis) was passedover the chip and the amount of binding recorded.

The table below show the results from cross-blocking assays on a rangeof different antibodies. The values in each square of the tablerepresent the amount of binding (in RU) seen when the antibodies (at 1.3μM on a binding site basis) or buffer indicated in the top row of thetable were mixed with the antibodies (at 1.3 uM on a binding site basis)or buffer indicated in the first column of the table.

Buffer Ab-4 Ab-13 Ab-A Ab-3 Ab-19 Buffer −0.5 693 428.5 707.3 316.1649.9 Ab-4 687.7 795.1 1018.2 860.5 869.3 822.5 Ab-13 425.6 1011.3 442.71108.4 431.9 1042.4 Ab-A 692.4 833.1 1080.4 738.5 946.2 868.1 Ab-3 305.5845.1 428.2 952.2 344.4 895.7 Ab-19 618.1 788.6 1022.5 863.3 891.5 658.7

Using the mean binding value (in RU) for each combination of antibodiesin the above table (since each combination appears twice) it is possibleto calculate the percentage of the theoretical binding shown by eachcombination of antibodies. The theoretical binding being calculated asthe sum of the average values for the components of each test mixturewhen assayed alone (i.e., antibody and buffer).

Buffer Ab-4 Ab-13 Ab-A Ab-3 Ab-19 Buffer Ab-4 90.75 60.45 85.4 60.75Ab-13 96.9 58.0 97.0 Ab-A 93.5 65.0 Ab-3 94.4 Ab-19

From the above data it is clear that Ab-4, Ab-A and Ab-19 cross-blockeach other. Similarly Ab-13 and Ab-3 cross block each other.

Example 7 ELISA-Based Cross-Blocking Assay

Liquid volumes used in this example would be those typically used in96-well plate ELISAs (e.g. 50-200 μl/well). Ab-X and Ab-Y, in thisexample are assumed to have molecular weights of about 145 Kd and tohave 2 sclerostin binding sites per antibody molecule. Ananti-sclerostin antibody (Ab-X) is coated (e.g. 50μ of 1 μg/ml) onto a96-well ELISA plate [e.g. Corning 96 Well EIA/RIA Flat Bottom Microplate(Product #3590), Corning Inc., Acton, Mass.] for at least one hour.After this coating step the antibody solution is removed, the plate iswashed once or twice with wash solution (e.g., PBS and 0.05% Tween 20)and is then blocked using an appropriate blocking solution (e.g., PBS,1% BSA, 1% goat serum and 0.5% Tween 20) and procedures known in theart. Blocking solution is then removed from the ELISA plate and a secondanti-sclerostin antibody (Ab-Y), which is being tested for it's abilityto cross-block the coated antibody, is added in excess (e.g. 50 μl of 10μg/ml) in blocking solution to the appropriate wells of the ELISA plate.Following this, a limited amount (e.g. 50 μl of 10 ng/ml) of sclerostinin blocking solution is then added to the appropriate wells and theplate is incubated for at least one hour at room temperature whileshaking. The plate is then washed 2-4 times with wash solution. Anappropriate amount of a sclerostin detection reagent [e.g., biotinylatedanti-sclerostin polyclonal antibody that has been pre-complexed with anappropriate amount of a streptavidin-horseradish peroxidase (HRP)conjugate] in blocking solution is added to the ELISA plate andincubated for at least one hour at room temperature. The plate is thenwashed at least 4 times with wash solution and is developed with anappropriate reagent [e.g. HRP substrates such as TMB (colorimetric) orvarious HRP luminescent substrates]. The background signal for the assayis defined as the signal obtained in wells with the coated antibody (inthis case Ab-X), second solution phase antibody (in this case Ab-Y),sclerostin buffer only (i.e. no sclerostin) and sclerostin detectionreagents. The positive control signal for the assay is defined as thesignal obtained in wells with the coated antibody (in this case Ab-X),second solution phase antibody buffer only (i.e. no second solutionphase antibody), sclerostin and sclerostin detection reagents. The ELISAassay needs to be run in such a manner so as to have the positivecontrol signal be at least 6 times the background signal.

To avoid any artifacts (e.g. significantly different affinities betweenAb-X and Ab-Y for sclerostin) resulting from the choice of whichantibody to use as the coating antibody and which to use as the second(competitor) antibody, the cross-blocking assay needs to be run in twoformats:

1) format 1 is where Ab-X is the antibody that is coated onto the ELISAplate and Ab-Y is the competitor antibody that is in solution

and

2) format 2 is where Ab-Y is the antibody that is coated onto the ELISAplate and Ab-X is the competitor antibody that is in solution.

Ab-X and Ab-Y are defined as cross-blocking if, either in format 1 or informat 2, the solution phase anti-sclerostin antibody is able to cause areduction of between 60% and 100%, specifically between 70% and 100%,and more specifically between 80% and 100%, of the sclerostin detectionsignal (i.e. the amount of sclerostin bound by the coated antibody) ascompared to the sclerostin detection signal obtained in the absence ofthe solution phase anti-sclerostin antibody (i.e. the positive controlwells).

In the event that a tagged version of sclerostin is used in the ELISA,such as a N-terminal His-tagged Sclerostin (R&D Systems, Minneapolis,Minn., USA; 2005 cat #1406-ST-025) then an appropriate type ofsclerostin detection reagent would include an HRP labeled anti-Hisantibody. In addition to using N-terminal His-tagged Sclerostin, onecould also use C-terminal His-tagged Sclerostin. Furthermore, variousother tags and tag binding protein combinations that are known in theart could be used in this ELISA-based cross-blocking assay (e.g., HA tagwith anti-HA antibodies; FLAG tag with anti-FLAG antibodies; biotin tagwith streptavidin).

Example 8 Cell Based Mineralization Assay for Identifying Agents Able toAntagonize Sclerostin Activity Introduction

Mineralization by osteoblast-lineage cells in culture, either primarycells or cell lines, is used as an in vitro model of bone formation.Mineralization takes from about one to six weeks to occur beginning withthe induction of osteoblast-lineage cell differentiation by one or moredifferentiation agents. The overall sequence of events involves cellproliferation, differentiation, extracellular matrix production, matrixmaturation and finally deposition of mineral, which refers tocrystallization and/or deposition of calcium phosphate. This sequence ofevents starting with cell proliferation and differentiation, and endingwith deposition of mineral is referred to herein as mineralization.Measurement of calcium (mineral) is the output of the assay.

Deposition of mineral has a strong biophysical characteristic, in thatonce mineral “seeds” begin to form, the total amount of mineral thatwill be deposited in the entire culture can sometimes be deposited quiterapidly, such as within a few days thereafter. The timing and extent ofmineral deposition in culture is influenced, in part, by the particularosteoblast-lineage cells/cell-line being used, the growth conditions,the choice of differentiation agents and the particular lot number ofserum used in the cell culture media. For osteoblast-lineagecell/cell-line mineralization cultures, at least eight to fifteen serumlots from more than one supplier should be tested in order to identify aparticular serum lot that allows for mineralization to take place.

MC3T3-E1 cells (Sudo H et al., In vitro differentiation andcalcification in a new clonal osteogenic cell line derived from newbornmouse calvaria. J. Cell Biol. 96:191-198) and subclones of the originalcell line can form mineral in culture upon growth in the presence ofdifferentiating agents. Such subclones include MC3T3-E1-BF (Smith E,Redman R, Logg C, Coetzee G, Kasahara N, Frenkel B. 2000.Glucocorticoids inhibit developmental stage-specific osteoblast cellcycle. J Biol Chem 275:19992-20001).

Identification of Sclerostin Neutralizing Antibodies

MC3T3-E1-BF cells were used for the mineralization assay. Ascorbic acidand B-glycerophosphate were used to induce MC3T3-E1-BF celldifferentiation leading to mineral deposition. The specific screeningprotocol, in 96-well format, involved plating cells on a Wednesday,followed by seven media changes (as described further below) over a12-day period with most of the mineral deposition taking place in thefinal approximately eighteen hours (e.g. Sunday night through Monday).For any given treatment, 3 wells were used (N=3). The specific timing,and extent, of mineral deposition may vary depending, in part, on theparticular serum lot number being used. Control experiments will allowsuch variables to be accounted for, as is well know in the art of cellculture experimentation generally.

In this assay system sclerostin inhibited one or more of the sequence ofevents leading up to and including mineral deposition (i.e., sclerostininhibited mineralization). Anti-sclerostin antibodies that were able toneutralize sclerostin's inhibitory activity allowed for mineralizationof the culture in the presence of sclerostin such that there was astatistically significant increase in deposition of calcium phosphate(measured as calcium) as compared to the amount of calcium measured inthe sclerostin-only (L e., no antibody) treatment group. For statisticalanalysis (using MS Excel and JMP) a 1-way-ANOVA followed by Dunnett'scomparison was used to determine differences between groups. Group meansfor each data set were considered significantly different when the Pvalue was less than 0.05 (P<0.05). A representative result from runningthis assay is shown in FIG. 22. In the absence of recombinant mousesclerostin, the sequence of events leading up to and including mineraldeposition proceeded normally. Calcium levels in each treatment groupare shown as means±Standard Error of the Mean (SEM). In this exemplaryexperiment calcium levels from the calcium assay were ˜31 μg/ml.However, addition of recombinant mouse sclerostin caused inhibition ofmineralization, and calcium was reduced by ˜85%. Addition ofanti-sclerostin monoclonal antibody Ab-19 or Ab-4 along with therecombinant sclerostin resulted in a statistically significant increasein mineral deposition, as compared to the sclerostin-only group, becausethe inhibitory activity of sclerostin was neutralized by eitherantibody. The results from this experiment indicate that Ab-19 and Ab-4are sclerostin neutralizing monoclonal antibodies (Mabs).

FIG. 23 shows a very similar result using recombinant human sclerostinand two humanized anti-sclerostin Mabs. FIG. 24 also shows a verysimilar result using recombinant human sclerostin and mouse andhumanized anti-sclerostin Mabs as indicated.

The antibodies used for the experiments shown in FIGS. 22, 23 and 24have molecular weights of about 145 Kd and have 2 sclerostin bindingsites per antibody molecule.

A detailed MC3T3-E1-BF cell culture protocol is described below.

Reagents and Medias

Reagents Company Catalog # Alpha-MEM Gibco-Invitrogen 12571-048 Ascorbicacid Sigma A4544 Beta-glycerophosphate Sigma G6376 100XPenStrepGlutamine Gibco-Invitrogen 10378-016 Dimethylsulphoxide (DMSO)Sigma D5879 or D2650 Fetal bovine serum (FBS) Cansera CS-008-500 (lot #SF50310) or Fetal bovine serum (FBS) TerraCell Int. CS-C08-1000A (lot #SF-20308)

Alpha-MEM is usually manufactured with a 1 year expiration date.Alpha-MEM that was not older than 6-months post-manufacture date wasused for the cell culture.

Expansion Medium (Alpha-MEM/10% FBS/PenStrepGlu) was prepared asfollows:A 500 ml bottle of FBS was thawed and filter sterilized through a 0.22micron filter.100 mls of this FBS was added to 1 liter of Alpha-MEM followed by theaddition of 10 mls of 100× PenStrepGlutamine. Unused FBS was aliquotedand refrozen for later use.Differentiation Medium (Alpha-MEM/10% FBS/PenStrepGlu, +50 μg/mlascorbic acid, +10 mM beta-glycerophosphate) was prepared as follows:100 mls of Differentiation Medium was prepared by supplementing 100 mlsof Expansion Medium with ascorbic acid and beta-glycerophosphate asfollows:

Stock cone (see below) Volume Final Conc. Ascorbic acid 10 mg/ml 0.5 mls100 μg/ml (50 ug/ml + 50 μg/ml) β-glycerophosphate 1M 1.0 mls 10 mM

Differentiation Medium was made by supplementing Expansion Medium onlyon the day that the Differentiation media was going to be used for cellculture. The final concentration of ascorbic acid in Differentiationmedium is 100 μg/ml because Alpha-MEM already contains 50 μg/ml ascorbicacid. Ascorbic acid stock solution (10 mg/ml) was made and aliquoted forfreezing at −80° C. Each aliquot was only used once (i.e. not refrozen).Beta-glycerophosphate stock solution (1 M) was made and aliquoted forfreezing at −20° C. Each aliquot was frozen and thawed a maximum of 5times before being discarded.

Cell Culture for Expansion of MC3T3-E1-BF Cells.

Cell culture was performed at 37° C. and 5% CO₂. A cell bank wasgenerated for the purposes of screening for sclerostin neutralizingantibodies. The cell bank was created as follows:

One vial of frozen MC3T3-E1-BF cells was thawed by agitation in a 37° C.water bath. The thawed cells were put into 10 mls of Expansion Medium(Alpha-MEM/10% FBS/PenStrepGlu) in a 50 ml tube and gently spun down for5 minutes. The cells were then resuspended in 4 mls of Alpha-MEM/10%FBS/PenStrepGlu. After determining the number of cells using trypan blueand hemacytometer, 1×10⁶ cells were plated in 50 mls Alpha-MEM/10%FBS/PenStrepGlu media in one T175 flask.

When this passage was confluent (at approximately 7 days), the cellswere trypsinized with trypsin/EDTA (0.05% Trypsin; 0.53 mM EDTA), gentlyspun down for 5 minutes and then resuspended in 5 mls Alpha-MEM/10%FBS/PenStrepGlu. After determining the number of cells using trypan blueand hemacytometer, cells were plated at 1×10⁶ cells in 50 misAlpha-MEM/10% FBS/PenStrepGlu media per one T175 flask. The number ofT175 flasks used for plating at this point depended upon the total cellnumber available and the desired number of flasks that were to be takenforward to the next passage. Extra cells were frozen down at 1-2×10⁶live cells/ml in 90% FBS/10% DMSO.

When this passage was confluent (about 3-4 days), the cells weretrypsinized with trypsin/EDTA (0.05% Trypsin; 0.53 mM EDTA), gently spundown for 5 minutes and then resuspended in 5 mls Alpha-MEM/10%FBS/PenStrepGlu. After determining the number of cells using trypan blueand hemacytometer, cells were plated at 1×10⁶ cells in 50 mlsAlpha-MEM/10% FBS/PenStrepGlu media per one T175 flask. The number ofT175 flasks used for plating at this point depended upon the total cellnumber available and the desired number of flasks that were to be takenforward to the next passage. Extra cells were frozen down at 1-2×10⁶live cells/ml in 90% FBS/10% DMSO.

When this passage was confluent (about 3-4 days), the cells weretrypsinized with trypsin/EDTA (0.05% Trypsin; 0.53 mM EDTA), gently spundown for 5 minutes and then resuspended in 5 mls Alpha-MEM/10%FBS/PenStrepGlu. After determining the number of cells using trypan blueand hemacytometer, cells were plated at 1×10⁶ cells in 50 mlsAlpha-MEM/10% FBS/PenStrepGlu media per one T175 flask. The number ofT175 flasks used for plating at this point depended upon the total cellnumber available and the desired number of flasks that were to be takenforward to the next passage. Extra cells were frozen down at 1-2×10⁶live cells/ml in 90% FBS/10% DMSO.

When this passage was confluent (about 3-4 days), the cells weretrypsinized with trypsin/EDTA (0.05% Trypsin; 0.53 mM EDTA), gently spundown for 5 minutes and then resuspended in 5 mls Alpha-MEM/10%FBS/PenStrepGlu. After determining the number of cells using trypan blueand hemacytometer, the cells were frozen down at 1-2×10⁶ live cells/mlin 90% FBS/10% DMSO. This “final passage” of frozen cells was thepassage that was used for the screening assay.

Cell Culture for Mineralizing MC3T3-E1-BF Cells.

Cell culture was performed at 37° C. and 5% CO₂. It is desirable tominimize temperature and % CO₂ fluctuations during the mineralizationcell culture procedure. This can be achieved by minimizing the time thatplates spend out of the incubator during feeding and also by minimizingthe number of times the incubator door is opened and closed during themineralization cell culture procedure. In this regard having a tissueculture incubator that is dedicated exclusively for the mineralizationcell culture (and thus not opened and closed more than is necessary) canbe helpful.

An appropriate number of “final passage” vials prepared as describedabove were thawed by agitation in a 37° C. water bath. The thawed cellswere put into 10 mls of Expansion Medium (Alpha-MEM/10% FBS/PenStrepGlu)in a 50 ml tube and gently spun down for 5 minutes. The cells were thenresuspended in 4 mls of Alpha-MEM/10% FBS/PenStrepGlu. After determiningthe number of cells by trypan blue and hemacytometer, 2500 cells wereplated in 200 microliters of Expansion media per well on collagen Icoated 96-well plates (Becton Dickinson Labware, cat #354407).

To avoid a mineralization plate-edge effect, cells were not plated inthe outermost row/column all the way around the plate. Instead 200microliters of PBS was added to these wells.

Exemplary Cell Culture Procedure

In the following procedure, the starting day for plating the cells isindicated to be a Wednesday. If a different day of the week is used asthe starting day for plating the cells, that day will trigger the dailyschedule for removing and adding media during the entire process asindicated below. For example, if the cells are plated on a Tuesday,media should not be removed and added on the first Friday and Saturday,nor on the second Friday and Saturday. With a Tuesday start, the plateswould be prepared for the calcium assay on the final Sunday.

Cells were plated on a Wednesday at 2500 cells in 200 μl of Expansionmedia.On Thursday all of the Expansion media was removed and 200 μl ofDifferentiation Media was added.On Friday 100 μl of media was removed and 100 μl of freshDifferentiation Media was added.On Monday 100 μl of media was removed and 100 μl of freshDifferentiation Media was added.On Tuesday 100 μl of media was removed and 100 μl of freshDifferentiation Media was added.On Wednesday 100 μl of media was removed and 100 μl of freshDifferentiation Media was added.On Thursday 100 μl of media was removed and 100 μl of freshDifferentiation Media was added.On Friday 100 μl of media was removed and 100 μl of freshDifferentiation Media was added.On the following Monday plates were prepared for the calcium assay asfollows:Plates were washed once with 10 mM Tris, HCl pH 7-8.Working under a fume hood, 200 μl of 0.5 N HCl was added per well.Plates were then frozen at −80° C.

Just prior to measuring calcium, the plates were freeze-thawed twice,and then trituration with a multichannel pipette was used to dispersethe contents of the plate. The contents of the plate was then allowed tosettle at 4° C. for 30 minutes at which point an appropriate amount ofsupernatant was removed for measuring calcium using a commerciallyavailable calcium kit. An exemplary and not-limiting kit is Calcium(CPC) Liquicolor, Cat. No. 0150-250, Stanbio Laboratory, Boerne, Tex.

In this cell based assay, sclerostin inhibits one or more of thesequence of events leading up to and including mineral deposition (i.e.sclerostin inhibits mineralization). Thus, in experiments wheresclerostin was included in the particular cell culture experiment, therecombinant sclerostin was added to the media starting on the firstThursday and every feeding day thereafter. In cases where ananti-sclerostin monoclonal antibody (Mab) was being tested for theability to neutralize sclerostin, i.e. allow for mineralization byneutralizing sclerostin's ability to inhibit mineralization, the Mab wasadded to the media starting on the first Thursday and every feeding daythereafter. According to the protocol, this was accomplished as follows:the Mab was preincubated with the recombinant sclerostin inDifferentiation media for 45-60 minutes at 37° C. and then this mediawas used for feeding the cells.

Described above is a 12-day mineralization protocol for MC3T3-E1-BFcells. Using the same reagents and feeding protocol, the originalMC3T3-E1 cells (Sudo H, Kodama H-A, Amagai Y, Yamamoto S, Kasai S. 1983.In vitro differentiation and calcification in a new clonal osteogeniccell line derived from newborn mouse calvaria. J Cell Biol 96:191-198)which we obtained from the RIKEN Cell Bank (RCB 1126, RIKEN BioResourceCenter 3-1-1 Koyadai, Tsukuba-shi, Ibaraki 305-0074 Japan) took longerto mineralize (20 days total for mineralization) than the MC3T3-E1-BFcells. Mineralization of the original MC3T3-E1 cells was inhibited byrecombinant sclerostin and this inhibition was blocked using asclerostin neutralizing antibody.

Example 9 Anti-Sclerostin Antibody Protects from Inflammation-InducedBone Loss in the CD4 CD45RB^(HI) Transfer Model of Colitis in SCID MiceSummary of Model

Injection of the CD45RB^(high) subset of CD4+ T cells into C.B-17 scidmice results in chronic intestinal inflammation with characteristicssimilar to those of human inflammatory bowel disease (IBD). Diarrhoeaand wasting disease is noted 3-5 weeks after cell transfer with severeleukocyte infiltration into the colon accompanied by epithelial cellhyperplasia and granuloma formation. C.B-17 scid mice which receive thereciprocal subset of CD4+ cells, those which express CD45RB^(low), donot exhibit colitis and have a weight gain indistinguishable fromuninjected scid mice. In addition to colitis symptoms, the CD4+CD45RB^(high) T cell transfer model of colitis is accompanied by areduction in bone mineral density (BMD), thought to be primarily throughinflammatory mechanisms rather than dietary malabsorption (Byrne, F. R.et al., Gut 54:78-86, 2005).

Induction of Colitis and Inflammation-Induced Bone Loss

Spleens were taken from female balb/c mice and disrupted through a 70 μmcell strainer. The CD4+ population was then enriched by negativeselection with Dynabeads using antibodies against B220, MAC-1, CD8 andI-A^(d). The enriched population was then stained with FITC conjugatedanti-CD4 and PE conjugated anti-CD45RB and fractionated intoCD4+CD45RB^(high) and CD4+CD45RB^(low) populations by two-color sortingon a Moflo (Dakocytomation). The CD45RB^(high) and CD45RB^(low)populations were defined as the brightest staining 40% and the dulleststaining 20% of CD4+ cells respectively. 5×10⁵ cells were then injectedi.p. into C.B-17 scid mice on day 0 and the development of colitis wasmonitored through the appearance of soft stools or diarrhoea and weightloss. Bone mineral density measurements were taken at the termination ofthe study (day 88).

Effect of Anti-Sclerostin Treatment on Colitis Symptoms and BMD

Ab-A IgG was dosed at 10 mg/kg s.c. from the day prior toCD4+CD45RB^(high) cell transfer and compared with mice which receivedthe negative control antibody 101.4 also dosed at 10 mg/kg s.c. Theantibodies were dosed weekly thereafter. A group of mice which receivednon-pathogenic CD4+CD45RB^(low) cells and were dosed with 10 mg/kg 101.4was studied as a control. At the termination of the study (day 88) thebone mineral density was measured and sections of the colon taken foranalysis of cell infiltration and assessment of histological damage.

a) No Effect on Colitis Symptoms

Typical colitis symptoms such as weight loss and infiltration ofinflammatory cells into the colon were unaffected by treatment withAb-A. Similarly there was no improvement of histological damage to thecolon after treatment with Ab-A.

b) Inhibition of Inflammation-Induced Loss of Bone Mineral Density.

On day 88 after transfer of cells into C.B-17 scid mice, the bonemineral density was measured (total BMD, vertebrae BMD and femur BMD).In comparison to control mice which received CD4+CD45RB^(low)non-pathogenic cells, mice which received CD4+CD45RB^(high) T cells andthe negative control antibody 101.4 had reduced bone mineral density, asshown in FIG. 25. In contrast, no reduction in BMD was noted aftertreatment with Ab-A. Total, vertebrae and femur measurements of BMD weresignificantly higher in mice receiving CD4+CD45RB^(high) T cells andtreated with Ab-A than mice receiving CD4+CD45RB^(high) T cells andtreated with 101.4 (P<0.001 by Bonferroni multiple comparison test).

Example 10 Kinexa-Based Determination of Affinity (K_(D)) ofAnti-Sclerostin Antibodies for Human Sclerostin

The affinity of several anti-sclerostin antibodies to human sclerostinwas assessed by a solution equilibrium binding analysis using KinExA®3000 (Sapidyne Instruments Inc., Boise, Id.). For these measurements,Reacti-Gel 6× beads (Pierce, Rockford, Ill.) were pre-coated with 40μg/ml human sclerostin in 50 mM Na2CO3, pH 9.6 at 4° C. overnight. Thebeads were then blocked with 1 mg/ml BSA in 1 M Tris-HCl, pH 7.5 at 4°C. for two hours. 10 pM, 30 pM, or 100 pM of the antibody was mixed withvarious concentrations of human sclerostin, ranging in concentrationfrom 0.1 pM to 1 nM, and equilibrated at room temperature for over 8hours in PBS with 0.1 mg/ml BSA and 0.005% P20. The mixtures were thenpassed over the human sclerostin coated beads. The amount of bead-boundanti-sclerostin antibody was quantified using fluorescent Cy5-labeledgoat anti-mouse-IgG or fluorescent Cy5-labeled goat anti-human-IgGantibodies (Jackson Immuno Research, West Grove, Pa.) for the mouse orhuman antibody samples, respectively. The amount of fluorescent signalmeasured was proportional to the concentration of free anti-sclerostinantibody in each reaction mixture at equilibrium. The dissociationequilibrium constant (K_(D)) was obtained from nonlinear regression ofthe competition curves using a n-curve one-site homogeneous bindingmodel provided in the KinExA Pro software. Results of the KinExA assaysfor the selected antibodies are summarized in the table below.

Antibodies Antigen K_(D) (pM) 95% confidence interval Ab-13 HumanSclerostin 0.6 0.4~0.8 pM   Ab-4  Human Sclerostin 3 1.8~4 pM Ab-19Human Sclerostin 3 1.7~4 pM Ab-14 Human Sclerostin 1 0.5~2 pM Ab-5 Human Sclerostin 6 4.3~8 pM Ab-23 Human Sclerostin 4 2.1~8 pM

Example 11 Biacore Method for Determining the Affinity of HumanisedAnti-Sclerostin Antibodies for Human Sclerostin

The BIAcore technology monitors the binding between biomolecules in realtime and without the requirement for labelling. One of the interactants,termed the ligand, is either immobilised directly or captured on theimmobilised surface while the other, termed the analyte, flows insolution over the captured surface. The sensor detects the change inmass on the sensor surface as the analyte binds to the ligand to form acomplex on the surface. This corresponds to the association process. Thedissociation process is monitored when the analyte is replaced bybuffer. In the affinity BIAcore assay, the ligand is the anti-sclerostinantibody and the analyte is sclerostin.

Instrument Biacore® 3000, Biacore AB, Uppsala, Sweden Sensor Chip

CM5 (research grade) Catalogue Number: BR-1001-14, Biacore AB, Uppsala,Sweden. Chips were stored at 4° C.

BIAnormalising Solution

70% (w/w) Glycerol. Part of BIAmaintenance Kit Catalogue Number:BR-1002-51, Biacore AB, Uppsala, Sweden. The BIAmaintenance kit wasstored at 4° C.

Amine Coupling Kit Catalogue Number: BR-1000-50, Biacore AB, Uppsala,Sweden.

Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). Madeup to 75 mg/mL in distilled water and stored in 200 μL aliquots at −70°C.N-Hydroxysuccinimide (NHS). Made up to 11.5 mg/mL in distilled water andstored in 200 μL aliquots at −70° C.1 M Ethanolamine hydrochloride-NaOH pH 8.5. Stored in 200 μL aliquots at−70° C.

Buffers

Running buffer for immobilising capture antibody: HBS-EP (being 0.01 MHEPES pH 7.4, 0.15 M NaCl, 3 mM EDTA, 0.005% Surfactant P20). CatalogueNumber: BR-1001-88, Biacore AB, Uppsala, Sweden. Buffer stored at 4° C.Immobilisation buffer: Acetate 5.0 (being 10 mM sodium acetate pH 5.0).Catalogue number: BR-1003-51, Biacore AB, Uppsala, Sweden. Buffer storedat 4° C.Running buffer for binding assay: HBS-EP (being 0.01 M HEPES pH 7.4,0.15 M NaCl, 3 mM EDTA, 0.005% Surfactant P20, Catalogue Number:BR-1001-88, Biacore AB, Uppsala, Sweden) with CM-Dextran added at 1mg/mL (Catalogue Number 27560, Fluka BioChemika, Buchs, Switzerland).Buffer stored at 4° C.

Ligand Capture

Affinipure F(ab′)₂ fragment goat anti-human IgG, Fc fragment specific.Jackson ImmunoResearch Inc (Pennsylvania, USA) Catalogue number:109-006-098. Reagent stored at 4° C.

Ligand

Humanised anti-human sclerostin antibodies Ab5, Ab14 and Ab20.

Analyte

Recombinant human sclerostin. Aliquots stored at −70° C. and thawed oncefor each assay.

Regeneration Solution

40 mM HCl prepared by dilution with distilled water from an 11.6 M stocksolution (BDH, Poole, England. Catalogue number: 101254H).5 mM NaOH prepared by dilution with distilled water from a 50 mM stocksolution. Catalogue number: BR-1003-58, Biacore AB, Uppsala, Sweden.

Assay Method

The assay format was capture of the anti-sclerostin antibody byimmobilised anti-human IgG-Fc then titration of the sclerostin over thecaptured surface.An example of the procedure is given below:BIA (Biamolecular Interaction Analysis) was performed using a BIAcore3000 (BIAcore AB). Affinipure F(ab′)₂ Fragment goat anti-human IgG, Fcfragment specific (Jackson ImmunoResearch) was immobilised on a CM5Sensor Chip via amine coupling chemistry to a capture level of ≈4000response units (RUs). HBS-EP buffer (10 mM HEPES pH 7.4, 0.15 M NaCl, 3mM EDTA, 0.005% Surfactant P20, BIAcore AB) containing 1 mg/mLCM-Dextran was used as the running buffer with a flow rate of 10 μl/min.A 10 μl injection of the anti-sclerostin antibody at −5 μg/mL was usedfor capture by the immobilised anti-human IgG-Fc. Antibody capturelevels were typically 100-200 RU. Sclerostin was titrated over thecaptured anti-sclerostin antibody at various concentrations at a flowrate of 30 μL/min. The surface was regenerated by two 10 μL injectionsof 40 mM HCl, followed by a 5 μL injection of 5 mM NaOH at a flowrate of10 μL/min.Background subtraction binding curves were analysed using theBIAevaluation software (version 3.2) following standard procedures.Kinetic parameters were determined from the fitting algorithm.The kinetic data and calculated dissociation constants are given inTable 2.

TABLE 2 Affinity of anti-sclerostin antibodies for sclerostin Antibodyka (1/Ms) kd (1/s) Kd (pM) Ab-5  1.78E+06 1.74E−04 97.8 Ab-14 3.30E+064.87E−06 1.48 Ab-20 2.62E+06 4.16E−05 15.8

Example 12 In Vivo Testing of Anti-Sclerostin Monoclonal Antibodies inCynomolgous Monkeys

Thirty-three, approximately 3-5 year old, female cynomolgus monkeys(Macaca fascicularis) were used in this 2-month study. The studycontained 11 groups:

Group 1: vehicle (N=4)Group 2: Ab-23 (N=2, dose 3 mg/kg)Group 3: Ab-23 (N=3, dose 10 mg/kg)Group 4: Ab-23 (N=3, dose 30 mg/kg)Group 5: Ab-5 (N=3, dose 3 mg/kg)Group 6: Ab-5 (N=3, dose 10 mg/kg)Group 7: Ab-5 (N=3, dose 30 mg/kg)Group 8: Ab-14 (N=3, dose 3 mg/kg)Group 9: Ab-14 (N=3, dose 10 mg/kg)Group 10: Ab-14 (N=3, dose 30 mg/kg)Group 11: Parathyroid Hormone (1-34) [PTH (1-34)] (N=3, dose 10 ug/kg)All dosing was subcutaneous. PTH (1-34) was dosed everyday, monoclonalantibodies (Mabs) were dosed twice (first dose at the beginning of thestudy and second dose at the one month time point). For assessment ofbone parameters (e.g. bone mineral density) pQCT (peripheralquantitative computed tomography) and DXA (dual energy X-rayabsorptiometry) scans were performed prior to the beginning of the study(to obtain baseline values) and after a month (prior to the second doseof Mab) and finally at the end of the study (2-month time point) atwhich point the monkeys were necropsied for further analysis (e.g.histomorphometric analysis). Animals were fluorochrome labeled (days 14,24, 47, and 57) for dynamic histomorphometry. Serum was collected atvarious time points during the study [day 1 pre-dose (the day of thefirst Mab dose), day 1 twelve hours post-dose, day 2, day 3, day 5, day7, day 14, day 21, day 28, day 29 twelve hours post-dose (day 29 was theday of the second and final Mab dose), day 30, day 31, day 33, day 35,day 42, day 49 and day 56].Three bone-related serum biomarkers were measured using commerciallyavailable kits:

Osteocalcin (OC) (DSL Osteocalcin Radioimmunoassay Kit; DiagnosticSystems Laboratories, Inc., Webster, Tex., USA) N-terminal Propeptide ofType I Procollagen (P1NP) (P1NP Radioimmunoassay Kit; Orion Diagnostica,Espoo, Finland)

C-telopeptide fragments of collagen type I al chains (sCTXI) (SerumCrossLaps® ELISA; Nordic Bioscience Diagnostics A/S, Herlev, Denmark).

pQCT and MCA scans yielded data on various bone parameters (includingbone mineral density (BMD) and bone mineral content) across numerousskeletal sites (including tibial metaphysis and diaphysis, radialmetaphysis and diaphysis, femur neck, lumbar vertebrae). Analysis ofthis bone data (percent change from baseline for each animal) and theanabolic (OC, P1NP) serum biomarker data (percent change from baselinefor each animal) revealed statistically significant increases, versusthe vehicle group, in some parameters at some of the time points anddoses for each Mab. This bone parameter data, serum biomarker data, aswell as the histomorphometric data, indicated that each of the 3 Mabs(Ab-23, Ab-5 and Ab-14) was able to neutralize sclerostin in cynomolgousmonkeys. This activity was most robust for Ab-23 and Ab-5, particularlyat the highest dose (30 mg/kg), with a clear increase in bone formation(anabolic effect) as well as net gains in bone (e.g. BMD). Statisticallysignificant increases in bone parameters and anabolic histomorphometricparameters were also found for the positive control group (PTH (1-34)).

Serum bone formation markers (P1NP, osteocalcin) were increased (p<0.05vs vehicle (VEH)) at various time points and doses, but particularly inthe 30 mg/kg groups for Ab-23 and Ab-5. Histomorphometric analysisrevealed dramatic increases (p<0.05 vs VEH) in bone formation rates incancellous bone at lumbar vertebra and proximal tibia (up to 5-foldincrease), as well as at the endocortical surface of the femur midshaft(up to 10-fold increase) at the higher doses of Ab-23 and Ab-5.Trabecular thickness was increased with high dose Ab-23 and Ab-5 inlumbar vertebrae (>60%, p<0.05 vs VEH). By study end (2 months), arealBMD, as percent change from baseline, was increased (p<0.05 vs VEH) atthe femur neck, ultra-distal radius (Ab-23, 30 mg/kg), and lumbarvertebrae (Ab-5, 30 mg/kg). The increases in areal BMD at the lumbarvertebrae were accompanied by increases in vertebral strength (97%increase in vertebral maximal load for Ab-23, 30 mg/kg; p<0.05 vs VEH);baseline values for lumbar areal BMD prior to Mab dosing werestatistically similar across all groups. In summary, short-termadministration of sclerostin-neutralizing Mabs in cynomolgous monkeysresulted, in part, in increases in bone formation, BMD and vertebralbone strength.

From the foregoing, although specific embodiments of the invention havebeen described herein for purposes of illustration, variousmodifications may be made without deviating from the spirit and scope ofthe invention. Accordingly, the invention is not limited except as bythe appended claims. All publications, published patent applications,and patent documents disclosed herein are hereby incorporated byreference.

What is claimed is:
 1. A sclerostin binding agent that cross-blocks thebinding of at least one of antibodies Ab-A, Ab-B, Ab-C, Ab-D, Ab-1,Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12,Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, Ab-22,Ab-23, and Ab-24 to sclerostin.
 2. The sclerostin binding agent of claim1 wherein said sclerostin binding agent is cross-blocked from binding tosclerostin by at least one of antibodies Ab-A, Ab-B, Ab-C, Ab-D, Ab-1,Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12,Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, Ab-22,Ab-23, and Ab-24.
 3. A sclerostin binding agent that is cross-blockedfrom binding to sclerostin by at least one of antibodies Ab-A, Ab-B,Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10,Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19, Ab-20,Ab-21, Ab-22, Ab-23, and Ab-24.
 4. The sclerostin binding agent of claim1 or 3 wherein the ability of said sclerostin binding agent tocross-block or to be cross-blocked is detected in a Biacore assay. 5.The sclerostin binding agent of claim 1 or 3 wherein the ability of saidsclerostin binding agent to cross-block or to be cross-blocked isdetected in an ELISA assay.
 6. The sclerostin binding agent of claim 1or 3 wherein said sclerostin binding agent is an antibody.
 7. Thesclerostin binding agent of claim 1 or 3 wherein said sclerostin bindingagent can increase at least one of bone formation, bone mineral density,bone mineral content, bone mass, bone quality and bone strength in amammal.
 8. The sclerostin binding agent of claim 1 or 3 wherein saidsclerostin binding agent can block the inhibitory effect of sclerostinin a cell based mineralization assay.
 9. A sclerostin binding agentwherein said sclerostin binding agent can block the inhibitory effect ofsclerostin in a cell based mineralization assay.
 10. A sclerostinbinding agent that binds to a Loop 2 epitope.
 11. A sclerostin bindingagent that binds to a T20.6 epitope.
 12. A sclerostin binding agent thatbinds to a “T20.6 derivative 1 (cystine-knot+4 arms)” epitope.
 13. Thesclerostin binding agent of any one of claims 7-12 wherein saidsclerostin binding agent is an antibody.
 14. A sclerostin binding agentthat comprises at least one CDR sequence having at least 75% identity toa CDR selected from SEQ ID NOs:39, 40, 41, 42, 43, 44, 45, 46, 47, 48,49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 78, 79, 80, 81,99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246,247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260,261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274,275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288,289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 351, 352, 353, 358,359, and
 360. 15. The sclerostin binding agent of claim 14 comprising atleast two of said CDR's.
 16. The sclerostin binding agent of claim 14comprising six of said CDR's.
 17. The sclerostin binding agent accordingto claim 14 wherein said percent identity is 85%.
 18. The sclerostinbinding agent according to claim 14 wherein said percent identity is95%.
 19. The sclerostin binding agent according to claim 14 comprising:a) CDR sequences of SEQ ID NOs:39, 40, and 41; b) CDR sequences of SEQID NOs:42, 43, and 44; c) CDR sequences of SEQ ID NOs:45, 46, and 47; d)CDR sequences of SEQ ID NOs:48, 49, and 50; e) CDR sequences of SEQ IDNOs:51, 52, and 53; f) CDR sequences of SEQ ID NOs:54, 55, and 56; g)CDR sequences of SEQ ID NOs:57, 58, and 59; h) CDR sequences of SEQ IDNOs:60, 61, and 62; i) CDR sequences of SEQ ID NOs:275, 276, and 277; j)CDR sequences of SEQ ID NOs:287, 288, and 289; k) CDR sequences of SEQID NOs:278, 279, and 280; l) CDR sequences of SEQ ID NOs:290, 291, and292; m) CDR sequences of SEQ ID NOs:78, 79, and 80; n) CDR sequences ofSEQ ID NOs:245, 246, and 247; o) CDR sequences of SEQ ID NOs:81, 99, and100; p) CDR sequences of SEQ ID NOs:248, 249, and 250 q) CDR sequencesof SEQ ID NOs:101, 102, and 103; r) CDR sequences of SEQ ID NOs:251,252, and 253; s) CDR sequences of SEQ ID NOs:104, 105, and 106 t) CDRsequences of SEQ ID NOs:254, 255, and 256; u) CDR sequences of SEQ IDNOs:107, 108, and 109 v) CDR sequences of SEQ ID NOs:257, 258, and 259w) CDR sequences of SEQ ID NOs:110, 111, and 112; x) CDR sequences ofSEQ ID NOs:260, 261, and 262; y) CDR sequences of SEQ ID NOs:281, 282,and 283; z) CDR sequences of SEQ ID NOs:293, 294, and 295; aa) CDRsequences of SEQ ID NOs:113, 114, and 115; bb) CDR sequences of SEQ IDNOs:263, 264, and 265; cc) CDR sequences of SEQ ID NOs:284, 285, and286; dd) CDR sequences of SEQ ID NOs:296, 297, and 298; ee) CDRsequences of SEQ ID NOs:116, 237, and 238; ff) CDR sequences of SEQ IDNOs:266, 267, and 268; gg) CDR sequences of SEQ ID NOs:239, 240, and241; hh) CDR sequences of SEQ ID NOs:269, 270, and 271; ii) CDRsequences of SEQ ID NOs:272, 273, and 274; jj) CDR sequences of SEQ IDNOs:242, 243, and 244; kk) CDR sequences of SEQ ID NOs:351, 352, and353; or ll) CDR sequences of SEQ ID NOs:358, 359, and
 360. 20. Thesclerostin binding agent according to claim 14 comprising: a) CDRsequences of SEQ ID NOs:54, 55, and 56 and CDR sequences of SEQ IDNOs:51, 52, and 53; b) CDR sequences of SEQ ID NOs:60, 61, and 62 andCDR sequences of SEQ ID NOs:57, 58, and 59; c) CDR sequences of SEQ IDNOs:48, 49, and 50 and CDR sequences of SEQ ID NOs:45, 46, and 47; d)CDR sequences of SEQ ID NOs:42, 43, and 44 and CDR sequences of SEQ IDNOs:39, 40, and 41; e) CDR sequences of SEQ ID NOs:275, 276, and 277 andCDR sequences of SEQ ID NOs:287, 288, and 289; f) CDR sequences of SEQID NOs:278, 279, and 280 and CDR sequences of SEQ ID NOs:290, 291, and292; g) CDR sequences of SEQ ID NOs:78, 79, and 80 and CDR sequences ofSEQ ID NOs: 245, 246, and 247; h) CDR sequences of SEQ ID NOs:81, 99,and 100 and CDR sequences of SEQ ID NOs:248, 249, and 250; i) CDRsequences of SEQ ID NOs:101, 102, and 103 and CDR sequences of SEQ IDNOs:251, 252, and 253; j) CDR sequences of SEQ ID NOs:104, 105, and 106and CDR sequences of SEQ ID NOs:254, 255, and 256; k) CDR sequences ofSEQ ID NOs:107, 108, and 109 and CDR sequences of SEQ ID NOs:257, 258,and 259; l) CDR sequences of SEQ ID NOs:110, 111, and 112 and CDRsequences of SEQ ID NOs:260, 261, and 262; m) CDR sequences of SEQ IDNOs:281, 282, and 283 and CDR sequences of SEQ ID NOs:293, 294, and 295;n) CDR sequences of SEQ ID NOs:113, 114, and 115 and CDR sequences ofSEQ ID NOs:263, 264, and 265; o) CDR sequences of SEQ ID NOs:284, 285,and 286 and CDR sequences of SEQ ID NOs:296, 297, and 298; p) CDRsequences of SEQ ID NOs:116, 237, and 238 and CDR sequences of SEQ IDNOs:266, 267, and 268; q) CDR sequences of SEQ ID NOs:239, 240, and 241and CDR sequences of SEQ ID NOs:269, 270, and 271; r) CDR sequences ofSEQ ID NOs:242, 243, and 244 and CDR sequences of SEQ ID NOs:272, 273,and 274; or s) CDR sequences of SEQ ID NOs:351, 352, and 353 and CDRsequences of SEQ ID NOs:358, 359, and
 360. 21. The sclerostin bindingagent of any one of claims 14-20 wherein said sclerostin binding agentis an antibody.
 22. A pharmaceutical composition comprising a sclerostinbinding agent according to any one of claims 1-12 and 14-20.
 23. Thecomposition of claim 22 wherein said sclerostin binding agent is anantibody.
 24. A sclerostin binding agent comprising at least one CDRsequence having at least 75% identity to a CDR selected from SEQ IDNOs:245, 246, 247, 78, 79, 80, 269, 270, 271, 239, 240 and
 241. 25. Asclerostin binding agent comprising at least one CDR sequence having atleast 75% identity to a CDR selected from CDR-H1, CDR-H2, CDR-H3,CDR-L1, CDR-L2 and CDR-L3 wherein CDR-H1 has the sequence given in SEQID NO:245 or SEQ ID NO:269, CDR-H2 has the sequence given in SEQ IDNO:246 or SEQ ID NO:270, CDR-H3 has the sequence given in SEQ ID NO:247or SEQ ID NO:271, CDR-L1 has the sequence given in SEQ ID NO:78 or SEQID NO:239, CDR-L2 has the sequence given in SEQ ID NO:79 or SEQ IDNO:240 and CDR-L3 has the sequence given in SEQ ID NO:80 or SEQ ID NO241.
 26. A sclerostin binding agent according to claim 25 comprisingthree CDRs, CDR-H1, CDR-H2 and CDR-H3 wherein (a) CDR-H1 is SEQ IDNO:245, CDR-H2 is SEQ ID NO:246 and CDR-H3 is SEQ ID NO:247 or (b)CDR-H1 is SEQ ID NO:269, CDR-H2 is SEQ ID NO:270 and CDR-H3 is SEQ IDNO:271.
 27. A sclerostin binding agent according to claim 25 comprisingthree CDRs, CDR-L1, CDR-L2 and CDR-L3 wherein (a) CDRL1 is SEQ ID NO:78,CDR-L2 is SEQ ID NO:79 and CDR-L3 is SEQ ID NO:80; or (b) CDRL1 is SEQID NO:239, CDR-L2 is SEQ ID NO:240 and CDR-L3 is SEQ ID NO:241.
 28. Asclerostin binding agent according to claim 25 comprising six CDRs,CDRH-1, CDR-H2, CDR-H3, CDR-L1 CDR-L2 and CDR-L3 wherein (a) CDR-H1 isSEQ ID NO:245, CDR-H2 is SEQ ID NO:246, CDR-H3 is SEQ ID NO:247, CDR-L1is SEQ ID NO:78, CDR-L2 is SEQ ID NO:79 and CDR-L3 is SEQ ID NO:80; or(b) CDR-H1 is SEQ ID NO:269, CDR-H2 is SEQ ID NO:270, CDR-H3 is SEQ IDNO:271, CDR-L1 is SEQ ID NO:239, CDR-L2 is SEQ ID NO:240 and CDR-L3 isSEQ ID NO:241.
 29. The sclerostin binding agent of any one of claims24-28 which is an antibody.
 30. The sclerostin binding agent of claim 29comprising a heavy chain wherein said heavy chain comprises apolypeptide having at least 85% identity to the sequence given in SEQ IDNO:333; SEQ ID NO:378; SEQ ID NO:327; SEQ ID NO:329; or SEQ ID NO:366.31. The sclerostin binding agent of claim 29 comprising a light chainwherein said light chain comprises a polypeptide having at least 85%identity to the sequence given in SEQ ID NO:332; SEQ ID NO:376; SEQ IDNO:314; SEQ ID NO:328; or SEQ ID NO:364.
 32. The sclerostin bindingagent of claim 29 comprising both a heavy chain and a light chainwherein (a) the heavy chain comprises a polypeptide having at least 85%identity to the sequence given in SEQ ID NO:333 and the light chaincomprises a polypeptide having at least 85% identity to the sequencegiven in SEQ ID NO:332; or (b) the heavy chain comprises a polypeptidehaving at least 85% identity to the sequence given in SEQ ID NO:378 andthe light chain comprises a polypeptide having at least 85% identity tothe sequence given in SEQ ID NO:376; or (c) the heavy chain comprises apolypeptide having at least 85% identity to the sequence given in SEQ IDNO:327 and the light chain comprises a polypeptide having at least 85%identity to the sequence given in SEQ ID NO:314; or (d) the heavy chaincomprises a polypeptide having at least 85% identity to the sequencegiven in SEQ ID NO:329 and the light chain comprises a polypeptidehaving at least 85% identity to the sequence given in SEQ ID NO:328; or(c) the heavy chain comprises a polypeptide having at least 85% identityto the sequence given in SEQ ID NO:366 and the light chain comprises apolypeptide having at least 85% identity to the sequence given in SEQ IDNO:364.
 33. The sclerostin binding agent of any one of claims 24-32which comprises a light chain and/or heavy chain constant region. 34.The sclerostin binding agent of claim 33 which comprises the IgG4 or theIgG2 constant region.
 35. A sclerostin binding agent having a heavychain comprising CDR's H1, H2 and H3 and comprising a polypeptide havingthe sequence provided in SEQ ID NO:137 or a variant thereof in whichsaid CDR's are at least 75% identical to SEQ ID NO:245, 246 and 247,respectively, and a light chain comprising CDR's L1, L2 and L3 andcomprising a polypeptide having the sequence provided in SEQ ID NO:133or a variant thereof in which said CDR's are at least 75% identical toSEQ ID NO:78, 79 and 80, respectively.
 36. A sclerostin binding agenthaving a heavy chain comprising CDR's H1, H2 and H3 and comprising apolypeptide having the sequence provided in SEQ ID NO:145 or 392 or avariant thereof in which said CDR's are at least 75% identical to SEQ IDNO:245, 246 and 247, respectively, and a light chain comprising CDR'sL1, L2 and L3 and comprising a polypeptide having the sequence providedin SEQ ID NO:141 or a variant thereof in which said CDR's are at least75% identical to SEQ ID NO:78, 79 and 80, respectively.
 37. A sclerostinbinding agent having a heavy chain comprising CDR's H1, H2 and H3 andcomprising a polypeptide having the sequence provided in SEQ ID NO:335or a variant thereof in which said CDR's are at least 75% identical toSEQ ID NO:269, 270 and 271, respectively, and a light chain comprisingCDR's L1, L2 and L3 and comprising a polypeptide having the sequenceprovided in SEQ ID NO:334 or a variant thereof in which said CDR's areat least 75% identical to SEQ ID NO:239, 240 and 241, respectively. 38.A sclerostin binding agent having a heavy chain comprising CDR's H1, H2and H3 and comprising a polypeptide having the sequence provided in SEQID NO:331 or a variant thereof in which said CDR's are at least 75%identical to SEQ ID NO:269, 270 and 271, respectively, and a light chaincomprising CDR's L1, L2 and L3 and comprising a polypeptide having thesequence provided in SEQ ID NO:330 or a variant thereof in which saidCDR's are at least 75% identical to SEQ ID NO:239, 240 and 241,respectively.
 39. A sclerostin binding agent having a heavy chaincomprising CDR's H1, 1-12 and H3 and comprising a polypeptide having thesequence provided in SEQ ID NO:345 or 396 or a variant thereof in whichsaid CDR's are at least 75% identical to SEQ ID NO:269, 270 and 271,respectively, and a light chain comprising CDR's L1, L2 and L3 andcomprising a polypeptide having the sequence provided in SEQ ID NO:341or a variant thereof in which said CDR's are at least 75% identical toSEQ ID NO:239, 240 and 241, respectively.
 40. A sclerostin binding agenthaving a heavy chain comprising a polypeptide having the sequenceprovided in SEQ ID NO:137, and a light chain comprising a polypeptidehaving the sequence provided in SEQ ID NO:133.
 41. A sclerostin bindingagent having a heavy chain comprising a polypeptide having the sequenceprovided in SEQ ID NO:145 or 392, and a light chain comprising apolypeptide having the sequence provided in SEQ ID NO:
 141. 42. Asclerostin binding agent having a heavy chain comprising a polypeptidehaving the sequence provided in SEQ ID NO:335, and a light chaincomprising a polypeptide having the sequence provided in SEQ ID NO:334.43. A sclerostin binding agent having a heavy chain comprising apolypeptide having the sequence provided in SEQ ID NO:331, and a lightchain comprising a polypeptide having the sequence provided in SEQ IDNO:330.
 44. A sclerostin binding agent having a heavy chain comprising apolypeptide having the sequence provided in SEQ ID NO:345 or 396, and alight chain comprising a polypeptide having the sequence provided in SEQID NO:341.
 45. A sclerostin binding agent according to any one of claims24-44 to which one or more effector or reporter molecule(s) is attached.46. An isolated polynucleotide sequence encoding the sclerostin bindingagent according to any one of claims 24-44.
 47. A cloning or expressionvector comprising one or more polynucleotide sequences according toclaim
 46. 48. A vector according to claim 47, wherein the vectorcomprises at least one sequence given in SEQ ID NO:134, 136, 138, 140,142, 144, 146, 148, 308, 310, 312, 342, 344, 346, 348, 349, 365, 367,373, 375, and
 379. 49. A host cell comprising one or more cloning orexpression vectors according to claim 47 or claim
 48. 50. A process forthe production of the sclerostin binding agent of any one of claims24-44, comprising culturing the host cell of claim 49 and isolating thesclerostin binding agent.
 51. A pharmaceutical composition comprising asclerostin binding agent according to any one of claims 24-45 incombination with one or more of a pharmaceutically acceptable excipient,diluent or carrier.
 52. A pharmaceutical composition according to claim51, additionally comprising other active ingredients.
 53. A sclerostinbinding agent according to any one of claims 24-45 or a pharmaceuticalcomposition according to claim 51 or claim 52, for use in the treatmentor prophylaxis of a pathological disorder that is mediated by sclerostinor that is associated with an increased level of sclerostin.
 54. Amethod for treating a bone-related disorder in a mammalian subject whichcomprises providing to a subject in need of such treatment apharmaceutical composition of claim
 22. 55. A method for treating abone-related disorder in a mammalian subject which comprises providingto a subject in need of such treatment a pharmaceutical composition ofclaim
 23. 56. A method for treating a bone-related disorder in amammalian subject which comprises providing to a subject in need of suchtreatment a pharmaceutical composition of claim
 51. 57. The methodaccording to claim 54, wherein the bone-related disorder is at least oneof achondroplasia, cleidocranial dysostosis, enchondromatosis, fibrousdysplasia, Gaucher's Disease, hypophosphatemic rickets, Marfan'ssyndrome, multiple hereditary exotoses, neurofibromatosis, osteogenesisimperfecta, osteopetrosis, osteopoikilosis, sclerotic lesions,pseudoarthrosis, pyogenic osteomyelitis, periodontal disease,anti-epileptic drug induced bone loss, primary and secondaryhyperparathyroidism, familial hyperparathyroidism syndromes,weightlessness induced bone loss, osteoporosis in men, postmenopausalbone loss, osteoarthritis, renal osteodystrophy, infiltrative disordersof bone, oral bone loss, osteonecrosis of the jaw, juvenile Paget'sdisease, melorheostosis, metabolic bone diseases, mastocytosis, sicklecell anemia/disease, organ transplant related bone loss, kidneytransplant related bone loss, systemic lupus erythematosus, ankylosingspondylitis, epilepsy, juvenile arthritides, thalassemia,mucopolysaccharidoses, Fabry Disease, Turner Syndrome, Down Syndrome,Klinefelter Syndrome, leprosy, Perthes' Disease, adolescent idiopathicscoliosis, infantile onset multi-system inflammatory disease, WinchesterSyndrome, Menkes Disease, Wilson's Disease, ischemic bone disease (suchas Legg-Calve-Perthes disease, regional migratory osteoporosis), anemicstates, conditions caused by steroids, glucocorticoid-induced bone loss,heparin-induced bone loss, bone marrow disorders, scurvy, malnutrition,calcium deficiency, osteoporosis, osteopenia, alcoholism, chronic liverdisease, postmenopausal state, chronic inflammatory conditions,rheumatoid arthritis, inflammatory bowel disease, ulcerative colitis,inflammatory colitis, Crohn's disease, oligomenorrhea, amenorrhea,pregnancy, diabetes mellitus, hyperthyroidism, thyroid disorders,parathyroid disorders, Cushing's disease, acromegaly, hypogonadism,immobilization or disuse, reflex sympathetic dystrophy syndrome,regional osteoporosis, osteomalacia, bone loss associated with jointreplacement, HIV associated bone loss, bone loss associated with loss ofgrowth hormone, bone loss associated with cystic fibrosis, chemotherapyassociated bone loss, tumor induced bone loss, cancer-related bone loss,hormone ablative bone loss, multiple myeloma, drug-induced bone loss,anorexia nervosa, disease associated facial bone loss, diseaseassociated cranial bone loss, disease associated bone loss of the jaw,disease associated bone loss of the skull, bone loss associated withaging, facial bone loss associated with aging, cranial bone lossassociated with aging, jaw bone loss associated with aging, and skullbone loss associated with aging and bone loss associated with spacetravel.
 58. The method according to claim 55, wherein the bone-relateddisorder is at least one of achondroplasia, cleidocranial dysostosis,enchondromatosis, fibrous dysplasia, Gaucher's Disease, hypophosphatemicrickets, Marfan's syndrome, multiple hereditary exotoses,neurofibromatosis, osteogenesis imperfecta, osteopetrosis,osteopoikilosis, sclerotic lesions, pseudoarthrosis, pyogenicosteomyelitis, periodontal disease, anti-epileptic drug induced boneloss, primary and secondary hyperparathyroidism, familialhyperparathyroidism syndromes, weightlessness induced bone loss,osteoporosis in men, postmenopausal bone loss, osteoarthritis, renalosteodystrophy, infiltrative disorders of bone, oral bone loss,osteonecrosis of the jaw, juvenile Paget's disease, melorheostosis,metabolic bone diseases, mastocytosis, sickle cell anemia/disease, organtransplant related bone loss, kidney transplant related bone loss,systemic lupus erythematosus, ankylosing spondylitis, epilepsy, juvenilearthritides, thalassemia, mucopolysaccharidoses, Fabry Disease, TurnerSyndrome, Down Syndrome, Klinefelter Syndrome, leprosy, Perthes'Disease, adolescent idiopathic scoliosis, infantile onset multi-systeminflammatory disease, Winchester Syndrome, Menkes Disease, Wilson'sDisease, ischemic bone disease (such as Legg-Calve-Perthes disease,regional migratory osteoporosis), anemic states, conditions caused bysteroids, glucocorticoid-induced bone loss, heparin-induced bone loss,bone marrow disorders, scurvy, malnutrition, calcium deficiency,osteoporosis, osteopenia, alcoholism, chronic liver disease,postmenopausal state, chronic inflammatory conditions, rheumatoidarthritis, inflammatory bowel disease, ulcerative colitis, inflammatorycolitis, Crohn's disease, oligomenorrhea, amenorrhea, pregnancy,diabetes mellitus, hyperthyroidism, thyroid disorders, parathyroiddisorders, Cushing's disease, acromegaly, hypogonadism, immobilizationor disuse, reflex sympathetic dystrophy syndrome, regional osteoporosis,osteomalacia, bone loss associated with joint replacement, HIVassociated bone loss, bone loss associated with loss of growth hormone,bone loss associated with cystic fibrosis, chemotherapy associated boneloss, tumor induced bone loss, cancer-related bone loss, hormoneablative bone loss, multiple myeloma, drug-induced bone loss, anorexianervosa, disease associated facial bone loss, disease associated cranialbone loss, disease associated bone loss of the jaw, disease associatedbone loss of the skull, bone loss associated with aging, facial boneloss associated with aging, cranial bone loss associated with aging, jawbone loss associated with aging, skull bone loss associated with aging,and bone loss associated with space travel.
 59. The method according toclaim 56, wherein the bone-related disorder is at least one ofachondroplasia, cleidocranial dysostosis, enchondromatosis, fibrousdysplasia, Gaucher's Disease, hypophosphatemic rickets, Marfan'ssyndrome, multiple hereditary exotoses, neurofibromatosis, osteogenesisimperfecta, osteopetrosis, osteopoikilosis, sclerotic lesions,pseudoarthrosis, pyogenic osteomyelitis, periodontal disease,anti-epileptic drug induced bone loss, primary and secondaryhyperparathyroidism, familial hyperparathyroidism syndromes,weightlessness induced bone loss, osteoporosis in men, postmenopausalbone loss, osteoarthritis, renal osteodystrophy, infiltrative disordersof bone, oral bone loss, osteonecrosis of the jaw, juvenile Paget'sdisease, melorheostosis, metabolic bone diseases, mastocytosis, sicklecell anemia/disease, organ transplant related bone loss, kidneytransplant related bone loss, systemic lupus erythematosus, ankylosingspondylitis, epilepsy, juvenile arthritides, thalassemia,mucopolysaccharidoses, Fabry Disease, Turner Syndrome, Down Syndrome,Klinefelter Syndrome, leprosy, Perthes' Disease, adolescent idiopathicscoliosis, infantile onset multi-system inflammatory disease, WinchesterSyndrome, Menkes Disease, Wilson's Disease, ischemic bone disease (suchas Legg-Calve-Perthes disease, regional migratory osteoporosis), anemicstates, conditions caused by steroids, glucocorticoid-induced bone loss,heparin-induced bone loss, bone marrow disorders, scurvy, malnutrition,calcium deficiency, osteoporosis, osteopenia, alcoholism, chronic liverdisease, postmenopausal state, chronic inflammatory conditions,rheumatoid arthritis, inflammatory bowel disease, ulcerative colitis,inflammatory colitis, Crohn's disease, oligomenorrhea, amenorrhea,pregnancy, diabetes mellitus, hyperthyroidism, thyroid disorders,parathyroid disorders, Cushing's disease, acromegaly, hypogonadism,immobilization or disuse, reflex sympathetic dystrophy syndrome,regional osteoporosis, osteomalacia, bone loss associated with jointreplacement, HIV associated bone loss, bone loss associated with loss ofgrowth hormone, bone loss associated with cystic fibrosis, chemotherapyassociated bone loss, tumor induced bone loss, cancer-related bone loss,hormone ablative bone loss, multiple myeloma, drug-induced bone loss,anorexia nervosa, disease associated facial bone loss, diseaseassociated cranial bone loss, disease associated bone loss of the jaw,disease associated bone loss of the skull, bone loss associated withaging, facial bone loss associated with aging, cranial bone lossassociated with aging, jaw bone loss associated with aging, skull boneloss associated with aging, and bone loss associated with space travel.60. A method of increasing at least one of bone formation, bone mineralcontent, bone mass, bone mineral density, bone quality, and bonestrength in a mammal comprising administering to the mammal apharmaceutical composition of claim
 22. 61. A method of increasing atleast one of bone formation, bone mineral content, bone mass, bonemineral density, bone quality, and bone strength in a mammal comprisingadministering to the mammal a pharmaceutical composition of claim 23.62. A method of increasing at least one of bone formation, bone mineralcontent, bone mass, bone mineral density, bone quality, and bonestrength in a mammal comprising administering to the mammal apharmaceutical composition of claim
 51. 63. A method of improving theoutcome in a mammal undergoing one or more of an orthopedic procedure,dental procedure, implant surgery, joint replacement, bone grafting,bone cosmetic surgery and bone repair such as fracture healing, nonunionhealing, delayed union healing and facial reconstruction, comprisingadministering to said mammal a pharmaceutical composition of claim 22before, during and/or after said procedure, replacement, graft, surgeryor repair.
 64. A method of improving the outcome in a mammal undergoingone or more of an orthopedic procedure, dental procedure, implantsurgery, joint replacement, bone grafting, bone cosmetic surgery andbone repair such as fracture healing, nonunion healing, delayed unionhealing and facial reconstruction, comprising administering to saidmammal a pharmaceutical composition of claim 23 before, during and/orafter said procedure, replacement, graft, surgery or repair.
 65. Amethod of improving the outcome in a mammal undergoing one or more of anorthopedic procedure, dental procedure, implant surgery, jointreplacement, bone grafting, bone cosmetic surgery and bone repair suchas fracture healing, nonunion healing, delayed union healing and facialreconstruction, comprising administering to said mammal a pharmaceuticalcomposition of claim 51 before, during and/or after said procedure,replacement, graft, surgery or repair.
 66. An antibody, wherein saidantibody is Ab-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6,Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16,Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, Ab-22, Ab-23, or Ab-24.
 67. Adiagnostic kit comprising a sclerostin binding agent according to anyone of claims 1, 3, 9-12 and 14-20.
 68. A diagnostic kit comprising anantibody according to claim
 6. 69. A diagnostic kit comprising anantibody according to claim
 21. 70. A diagnostic kit comprising anantibody according to claim
 29. 71. A polypeptide comprising at leastone of SEQ ID NOs:39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51,52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 78, 79, 80, 81, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248,249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262,263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276,277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290,291, 292, 293, 294, 295, 296, 297, 298, 351, 352, 353, 358, 359, and360.
 72. A polypeptide according to claim 71 conjugated to at least oneof Fc, PEG, albumin, and transferrin.